Laboratory Manual for Biomedical Research

Laboratory Manual for Biomedical Research

New Edition

Chapter 1. General Lab Techniques

Lab security and basic techniques
Advanced lab skills
Advanced lab skills (2)

Chapter 2. Molecular Biology -- Molecular Separation

Molecular Separation
DNA Agarose Gel Electrophoresis
RNA Agarose Gel Electrophoresis
SDS-Page - Polyacrylamide gel electrophoresis
2D Page-Two - Dimensional Polyacrylamide Gel Electrophoresis
Ion Exchange Chromatography
Gel Filtration Chromatography
Affinity chromatography

Chapter 3. Molecular Biology -- DNA and RNA

Nucleic acid methods (1)
Nucleic acid methods (2)
DNA isolation & related protocols
DNA Purification (glass milk vs electroelution)
DNA, RNA Sequencing
RNA Isolation and Purification
Isolation of DNA,RNA, and Protein simultaneously.
DNA mutation detection by SSCP
Preparation of DNA and RNA probes
Southern blot hybridization
Northern blot hybridization
Loss of Heterozygosity (LOH)
Gene knockout protocol
SiRNA gene knockout
Plasmid and its usefulness
DNA library construction
Microarray protocols.
Basic knowledge of microarray.
Introduction to Microarray.
MicroArray Procedure
Total RNA Isolation from cultured cells.
DNase Treatment of Total RNA
Making the single strand cDNA probe.
Automated Slide Processor (ASP) Version for hybridization.
Washing microarrays in ASP.
Processing of Array slide
Pre-hybridization of the processed slides (NON-Automated version).
Hybridization of Cy3 + Cy5 probe to glass array (NON-Automated version).
Preparation of Dendrimer Cy3 and Cy5.
Washing unbound probe from glass array (NON-Automated version).
Hybridization of Dendrimers (Cy3 and Cy5) to Array (NON-Automated version).
Washing unbound dendrimer from glass array(NON-Automated version).
Microarray Dababases
Troubleshooting
Other Microarray Protocols (1,2)
Gene transfection
Gene therapy for cancer
Molecular cloning
Conditional gene transfection(Tet on/off)

Chapter 4. Genetics

Epigenetics protocols
Mutagenesis protocols
Single nucleotide polymorphisms (SNPs)

Chapter 5. Molecular Biology --PCR Serials

PCR,RT-PCR,Real time PCR etc.
PCR (General Procedure)
Recommended Reagent Concentrations
Recommended Reaction Conditions
Initial Conditions
Temperature Cycling
"Hot Start" PCR
Asymmetric PCR for ssDNA Production
Detecting Products
Labelling PCR Products with Digoxigenin
Cleaning PCR Products
Sequencing PCR Products
Cloning PCR Products
AND ALWAYS REMEMBER
PCR Primer Design Tools
RT-PCR
Real time PCR
More PCR Protocols Online
Video and Animation of PCR
Mouse Genotyping by PCR
PCR Based Molecular Cloning
PCR Troubleshootings

Chapter 6. Molecular Biology – Protein

Protein methods
Protein labeling techniques
Protein sequencing
Subcellular fractionations
Western blot hybridization
Western Blot Hybridization Center
Western Blot Full Procedure (from Pierce)
Western Blotting Procedure with Video Show
Protocol 1: Western Blot
Sample protein preparation
Protein concentration assay
Electrophoresis and blotting
Blocking non-specific antigen
Incubation with primary antibody
Incubation with secondary antibody
Substrate (ECL) incubation
Restore a western blot using pierce stripping buffer
Western blot related solutions and buffers
Western blot handbook
Western blot troubleshooting
Protocol 2: Western Blot 2.
Protocol 3: Western Blot 3.
More online Western blot protocols (1) (2)
Protein chips
Kinase assay
Methods for detecting protein phosphorylation

Chapter 7. DNA Protein Interactions

EMSA
ChIP assay
Filter Binding (1) (2)
DNase Footprinting (1) (2)
DMS Footprinting
Genetic analysis
X-ray crystallography
Methylation Interference
Mapping protien/DNA interaction by cross-linking

Chapter 8. Immunohistochemistry/immunology

Immunology/Histology
Preparing silanized (+plus) slides
Microscopy Techniques
Confocal microscope technique
Electron microscopy
HE staining
DAPI & PI nucleic acid stain
Hybridization in situ
FISH
Special cell & cell fraction stains
Antibody purification
Antibody storage and handling
Conjugation of monoclonal antibodies
Antigen retrieval
Immunoperoxidase staining techniques
Immunofluorohistochemistry
Immunoprecipitation
Histotechnology--technical methods
FRQs for histochemistry
Laser Capture Microdissection
Elisa

Chapter 9. Cellular Biology

General cell culture protocols
Chromosome karyotype
Proliferation assays (MTT, BrdU, 3H-Thymidine incoporation)
Cell cycle assay
Migration assay
Stem cell & related protocols
Introduction
What are the unique properties of all stem cells?
What are embryonic stem cells?
What are adult stem cells?
What are the similarities and differences between embryonic and adult stem cells?
What are the potential uses of human stem cells and the obstacles that must be overcome before these potential uses will be realized?
Where can I get more information?
International Society for Stem Cell Research (ISSCR)
Culture of Human Embryonic Stem Cells (hESC)
Establishment and characterization of human embryonic stem cell line
Rat stem cell isolation and culture
Mouse stem cell isolation and culture
Induction of Stem Cell Differentiation
Cancer stem cells
Video Data on Stem Cell Research (click to show)
Apoptosis and related protocols
Soft Agar Assay for Colony Formation
Aorta ring assay
GFP transfection
Blood cell fractionation (white blood cell isolation)
Endothelial cell isolation and culture protocols
Flow cytometry (FCM)

Chapter 10. Metabolism, GC/MS, NMR and Proteomics

GC/MS Background
Glucose metabolism and its related protocols
PAS staining
Deoxyribose procedure
Ribose metabolism analysis
Polysaccharide sequencing
Lactate cleanup and derivative
Amino acids
Fatty acids
Lipid protocols
Cholesterol
Bile acids
Urea procedure
Choline Incorporation Assay
Isotope Ratio Mass Spectrometer (IRMS)
Liquid chromatography / mass spectrometer (LC/MS)
Proteomics
NMR protocols and tutorials

Chapter 11. Animal Experiments

Blood sampling from animals
Basic skills for animal experiments
Anatomy and physiology of animals
Cancer xenograft animal models
Transgenic animal procedures
Transgenic cancer models
Animal models for depression-like and anxiety-like behavior

Chapter 12. Worm: C. Elegans

C. Elegans Protocols

Chapter 13. HPLC and TLC

HPLC protocols
TLC

Chapter 14. Buffers and Solutions in Lab.

Commonly Used Buffer and solution (114 formats in alphabeta)
Compound solutions (Practical Molecular Biology)How to Make Simple Solutions and Dilutions (Department of Biology, Bates College)Reto's Buffers&Media Book (Private Page)Salt solutions(Practical Molecular Biology)Single Component Solutions (Practical Molecular Biology)Buffers (Gerard R. Lazo)
Antibiotics (Gerard R. Lazo)Buffer Listing (Salmon Lab.)
Cell Culture Media and Solutions (Donis-Keller lab)
Commonly used solutions (1)--- 1 ) 2 ) 3 ) 4 ) 5 ) 6 )
Commonly used solutions (2)--- 1 ) 2 ) 3 )
Solutions for molecular cloning---1 ) 2 ) 3 ) 4 ) 5 ) 6 ) 7 ) 8 ) 9 )

Chapter 15. Other Resources

Free eBooks at Library Online
Cinema Online, Free Movies --(1) (2) (3)
Progresses in Life Science
Progress in Cancer Apoptosis -- (1) (2) (3)
Progress in Gene Therapy --- (1) (2) (3)
Progress in Neuronal Regeneration -- (1) (2) (3)
Progress in AIDS Treatment -- (1) (2) (3)
Progress in Organ Transplantation -- (1) (2) (3)
Progress in Stem Cell Research -- (1) (2) (3)
Free eBooks in life science (1)
Free eBooks in life science (2,3)
Most Updated Biomedical Books
Pathway databases
Pathway Search
Biomedical Job Opportunities
Tools for Statistics
Free Software pDRAW32 to Draw DNA Analysis Charts
Biological Educational Resources
Genetics Education
Resources of Medical Biochemistry
Textbooks and Lab Manuals

Copyright: 2008-2009 (c) www.Lab-Manual.Com All Rights Reserved

Contact US: labmanual.com@gmail.com

Stem Cells, Regenerative Medicine, and Animal Models of Disease

Dennis A. Steindler

Abstract

The field of stem cell biology and regenerative medicine is rapidly moving toward translation to clinical practice, and in doing so has become even more dependent on animal donors and hosts for generating cellular reagents and assaying their potential therapeutic efficacy in models of human disease. Advances in cell culture technologies have revealed a remarkable plasticity of stem cells from embryonic and adult tissues, and transplantation models are now needed to test the ability of these cells to protect at-risk cells and replace cells lost to injury or disease. With such a mandate, issues related to acceptable sources and controversial (e.g., chimeric) models have challenged the field to provide justification of their potential efficacy before the passage of new restrictions that may curb anticipated breakthroughs. Progress from the use of both in vitro and in vivo regenerative medicine models already offers hope both for the facilitation of stem cell phenotyping in recursive gene expression profile models and for the use of stem cells as powerful new therapeutic reagents for cancer, stroke, Parkinson's, and other challenging human diseases that result in movement disorders. This article describes research in support of the following three objectives: (1) To discover the best stem or progenitor cell in vitro protocols for isolating, expanding, and priming these cells to facilitate their massive propagation into just the right type of neuronal precursor cell for protection or replacement protocols for brain injury or disease, including those that affect movement such as Parkinson's disease and stroke; (2) To discover biogenic factors—compounds that affect stem/progenitor cells (e.g., from high-throughput screening and other bioassay approaches)—that will encourage reactive cell genesis, survival, selected differentiation, and restoration of connectivity in central nervous system movement and other disorders; and (3) To establish the best animal models of human disease and injury, using both small and large animals, for testing new regenerative medicine therapeutics.

Key Words: drug discovery; human therapeutics; recursive gene profiling; regenerative medicine; stem cell; transplantation

Introduction

Stem cells have the propensity to produce tissue, an attribute that not only contributes to normal human development but also can lead to oncogenic transformation and hyperplasia (Gibbs et al. 2005; Ignatova et al. 2002; Steindler 2006). Two characteristics of stem (and progenitor) cells reveal their dual nature: (1) "poiesis" (generation) and (2) the overgeneration of cells and tissue (the so-called oncogenic transformation that leads to neoplasia). Because of these particular attributes, there is widespread interest in stem cells and regenerative medicine and their potential to treat and cure human diseases. But controversy and debates surround the question of which cells might be both the best and the most ethically acceptable therapeutic reagents, likewise determining which animal models are indeed the most effective. Animal models of disease are certainly necessary for the regenerative medicine field. Clinical trials of adult (e.g., bone marrow or cord blood transplantation) as well as fetal stem or progenitor cells have already demonstrated the efficacy of such regenerative medicine cell therapies for protecting, repairing, and replacing at-risk cells and tissues (Bjorklund 2005; Reier 2004; Press Release, Yahoo! Finance, November 15, 2006). Yet there is also a daunting side to the emerging field of regenerative medicine. Great expectations and desperate hope for immediate clinical application have driven intense debates at the state level, and international hearings to establish guidelines also try to respond to demands from different citizen groups with disparate agendas. The situation has also prompted patients all over the world to seek alternative and usually unproven stem cell "therapies" that can put them at risk. These challenges justify support for more science that must include both in vitro and in vivo studies of stem/progenitor cells from a variety of tissues and organs.

This article describes advances to date in the use of cells and animal models in regenerative medicine, expectations for future discoveries of the best stem and progenitor cell populations from different tissues and organs, and how in vitro high-throughput screening (HTS¹) bioassays might best utilize the potency of embryonic, fetal, and adult stem cells. The article also describes uses of modeling, in vitro studies, and dynamic stem cell and biogenic stem cell factor screening that could lead to more rapid developments in translational regenerative medicine. The reasoning below suggests that research in regenerative biology and regenerative medicine, although human-centric because of the eventual need for cells from a variety of human tissues and organs at different stages of development and aging, nonetheless requires animals, both as sources of immature cells and as recipients for cell and engineered tissue grafts to establish therapeutic proof-of-principle for any new cell or drug therapy. In vitro bioassay screening and use of simpler organisms could reduce the need for experimentation with mammalian models once researchers better understand the nature of different stem/progenitor cell populations and also further refine HTS. Thus, it is worthwhile to further develop cell culture assays, explore virtual gene and protein screens, and establish standardized and efficient rodent and other animal models of human disease to generate universal bioassays that can be used to establish the required safety and efficacy of any potential new regenerative medicine therapy before going on to human clinical trials. In particular, immunocompromised animals with diseased and injured tissues should continue to host human cell transplants, and investigators should continue to test new drugs gleaned from studies of the bioactive compounds associated with the growth and differentiation of stem cells in the same animal models.

There is no question that animal models of stem cell research in support of regenerative medicine will facilitate rapid translation to the bedside. The regenerative medicine field will continue to foster respect for the animal kingdom amid a pressing need to find new cures for human suffering. With the remarkable paradigm shift that has occurred in scientists' understanding of human self-regenerative potential, there is a high level of confidence that stem cell biology and regenerative medicine will lead to exceptionally effective new therapeutics for movement disorders and all other neurological challenges in the not too distant future.

……

The full article is available via below link.

Stem cells, regenerative medicine, and animal models of disease.

Stem cells, regenerative medicine, and animal models of disease. Steindler DA. Program in Stem Cell Biology and Regenerative Medicine, University of Florida ... www.ncbi.nlm.nih.gov/pubmed/17712220

The world of stem cells

The world of stem cells
A newly fertilized egg has cells that have no particular function.
Stem cells from embryos can become any kind of cell in the human body.
We are aware that different types of cells make up our body (e.g., blood cells, skin cells, cervical cells) but usually forget to appreciate that all of these different cell types arose from a single cell, the fertilised egg. Developmental biologists study the awesome events that occur between the fertilised egg and the formation of a new individual.
 The first steps simply involve cell division: one cell becomes two cells; two cells become four cells, etc.
 Each of these individual cells of early development is not specialized (undifferentiated), that is it does not have a specific body function, and has the capability to contribute to all of the organs in an individual and thus are called totipotent.
 These cells are embryonic stem (ES) cells and have both the capacity to self-renew, thus maintaining a continuous supply of stem cells and the ability to give rise to specialized (differentiated) cell types, such as liver cells or brain cells.
 It is believed that once differentiated, cells remain so and usually lose their ability to divide.
Stem cells from adults can also be used in cell therapy, with limitations.
Stem cells also exist in adults and allow specific tissues to regenerate throughout life. They also have the ability for self-renewal and multi-lineage differentiation. In fact, the list for identifying adult stem cells and lineage specific progenitor cells (with limited self-renewal ability) is growing.
Sources of stem cells
The main clinical application of stem cells is as a source of donor cells to be used to replace cells in transplantation therapy. Stem cells can be obtained from several sources:
 Spare embryos: stem cells can come from leftover embryos stored at fertility clinics that were not used by couples to have children.
 Special purpose embryos: embryos are created in vitro fertilization (artificially in the lab) for the sole purpose of extracting their stem cells.
Embryos and living or dead adult tissue provide stem cells.
 Cloned embryos: embryos are cloned in labs using somatic nuclear transfer method in order to harvest their stem cells.
 Aborted fetuses: stem cells are taken from fetuses in early development that have been aborted.
 Umbilical cords: this after-childbirth tissue holds potential for research.
 Adult tissue or organs: stem cells are obtained from the tissue or organs of living adults during surgery.
 Cadavers: isolation and survival of neural progenitor cells from human post-mortem tissues (up to 20 hours after death) has been reported and provides an additional source of human stem cells.1
Embryonic stem cells must be obtained when an embryo is in early development, that is, when the fertilised egg has divided to form about 1000 cells. These cells are separated and maintained in a cell culture dish, thereby halting embryonic development towards creating an individual. This is why embryonic stem cell research is the subject of ethical debates. Utilization of adult stem cells pose less of an ethical dilemma: however, adult stem cells may not have the same potential as those derived from embryos for medical therapeutics.
Comparing embryonic and adult stem cells
Embryonic stem cells have advantages and disadvantages for therapy.
Advantages: They are
Embryos can contribute an endless supply of stem cells.
 Flexible: They have the potential to make any body cell.
 Immortal: One cell line can potentially supply endless amounts of cells with carefully defined characteristics.
 Easily available: human embryos can be obtained from fertility clinics.
Disadvantages: They could be
 Difficult to control: The method for inducing the cell type needed to treat a particular disease must be defined and optimized .
 At odds with a patient’s immune system: It is possible that transplanted cells would differ in their immune profile from that of the recipient and so would be rejected.
 Ethically controversial: Those who believe life begins at conception say that doing research on human embryos is unethical even if donors give their consent.
Adult stem cells also have good and difficult characteristics for therapy.
Advantages: They are
 Already somewhat specialized: Inducement may be simpler.
 Immune hardy: Recipients who receive the products of their own stem cells will not experience immune rejection.
 Flexible: Adult stem cells may be used to form other tissue types.
 Mixed degree of availability: Some adult stem cells are easy to harvest and others, such as neural (brain) stem cells, can be dangerous to the donor.
Adult stem cells are sometimes hard to obtain and don’t last long.
Disadvantages: They could be
 Minimal quantity: They are difficult to obtain in large quantities.
 Finite: They don’t live as long in a culture as embryonic stem cells.
 Genetically unsuitable: The harvested stem cells may carry genetic mutations for disease or become defective during experimentation.
Stem cells can develop into liver, heart, blood, or any other cell.
The surprising property of adult stem cells: transdifferentiation
Adult stem cells were thought to be restricted to produce differentiated cells, which were specific to the organ from which they were isolated. Recently, several examples have been reported which demonstrate that these stem cells, under certain conditions, can be induced to form other cell types (transdifferentiation). For example:
 neural stem cells (NSC) can give rise to blood and skeletal muscle
 bone marrow cells can give rise to muscle, liver cells, and astrocytes
Stem cells can be transplanted directly into the patient.
When NSCs were used to form muscle, no inducers were needed other than co-culturing them with muscle progenitor cells (myoblasts) or injecting them into muscle.2 This holds promise for cell transplantation therapies in that the experiment suggests that host tissue can instruct transplanted cells to a desired result. Scientists, then, can consider whether it is best for stem cells to be differentiated in vitro (artificially) prior to transplantation or by transplanting them directly into the defective tissue. Some experiments have shown that naturally transplanted stem cells were able to migrate to regions where cells had died due to stroke (called ischaemia).
Stem cell therapies
Stem cells can renew blood and bones after chemotherapy.
Stem cells offer the opportunity of transplanting a live source for self-regeneration. Bone marrow transplants (BMT) are a well known clinical application of stem cell transplantation. BMT can repopulate the marrow and restore all the different cell types of the blood after high doses of chemotherapy and/or radiotherapy, our main defence used to eliminate endogenous cancer cells. The isolation of additional stem and progenitors cells is now being developed for many other clinical applications. Several are described below.
Stem cells from hair can grow into skin.
Skin replacement The knowledge of stem cells has made it possible for scientists to grow skin from a patient’s plucked hair. Skin (keratinocyte) stem cells reside in the hair follicle and can be removed when a hair is plucked.3 These cells can be cultured to form an epidermal equivalent of the patients own skin and provides tissue for an autologous graft, bypassing the problem of rejection. It is presently being studied in clinical trials as an alternative to surgical grafts used for venous ulcers and burn victims.
Brain cell transplantation Neural stem cells were only until recently thought to be strictly embryonic. Many findings have proved this incorrect. The identification and localisation of neural stem cells, both embryonic and adult, has been a major focus of current research. Potential targets of neural stem cell transplants include stroke, spinal cord injury, and neurodegenerative diseases such as Parkinson’s Disease.
Stem cells can provide dopamine - a chemical lacking in victims of Parkinson’s Disease.
Parkinson’s Disease involves the loss of cells which produce the neurotransmitter dopamine. The first double-blind study of fetal cell transplants for Parkinson’s Disease reported survival and release of dopamine from the transplanted cells and a functional improvement of clinical symptoms.4 However, some patients developed side effects, which suggested that there was an oversensitization to or too much dopamine. Although the unwanted side effects were not anticipated, the success of the experiment at the cellular level is significant. Again, further studies are needed and ongoing. Over 250 patients have already been transplanted with human fetal tissue.
Several biotechnology companies are developing different strategies of stem cell therapies.
 Diacrin has been developing xenotransplants using fetal pig cells. Clinical trials for chronic stroke patients have begun. Presently, stroke patients require treatment within 24 hours after stroke for effective therapeutic results. Many patients do not receive treatment in time because the symptoms are not initially obvious. Diacrin’s therapy could be applied weeks to months after the initial trauma.
 NeuroNova’s strategy is to culture adult human cells from donors, differentiate them in culture to produce the cell type (dopaminergic neurons) which is lost in Parkinson Disease, and to transplant them into the brain of patients.
 Neurotech is using genetically altered brain endothelial cells (engineered to produce human Interleukin-2) as immunotherapy for gliomas. Results from experiments in rats showed that these cells "mopped up" the tumour cells and as a result a clinical study has commenced.
Mouse stem cells were made to produce their own insulin.
Treatment for diabetes Diabetes affects 16 million people in the U.S. and is caused by the abnormal metabolism of insulin. Normally, insulin is produced and secreted by the cellular structures called the islets of Langerhans in the pancreas. Recently, insulin expressing cells from mouse stem cells have been generated.5 In addition, the cells self assemble to form structures, which closely resemble normal pancreatic islets and produce insulin. Future research will need to investigate how to optimise conditions for insulin production with the aim of providing a stem cell-based therapy to treat diabetes to replace the constant need for insulin injections.
Future directions
Mouse brain stem cells could self-repair.
The generation of new neurons in the adult brain is limited. However, self-repair of neuronal cell death has been recently demonstrated in the mouse and suggests that stem cells which normally reside in the brain may someday be able to be stimulated by inducers in a manner similar to how we induce our immune system by vaccination.6 This would bypass the need for cell transplantation. Intensive research needs to be pursued into the cell mechanisms involved.
The potential of embryonic stem cells to provide other differentiated cell types needs to be investigated. The production of cardiac muscle cells, which have thus far been evasive, would hold tremendous promise for the number one killer: heart disease.
Scientists and stem cell research
Poll: The majority of Americans favor stem cell research.
Scientists believe that stem-cell research could lead to cures for a myriad of diseases afflicting humans. Anti-abortion groups, some religious groups, and conservative citizens say that using cells from embryos is immoral because it destroys life. However, a recent ABCNews/Beliefnet poll has shown that Americans support stem cell research by a 2-1 margin and say that it should be funded by the federal government, despite controversy over the use of human embryos.7
Conclusion: Stem cell research should be pursued but under legislative guidance.
Most scientists Do Not support applications for human reproductive cloning (that is, they do not want any embryos altered during stem cell research to develop past a defined stage). They agree with governments and concerned citizens that it should be banned worldwide. However, they Do want the opportunity to continue stem cell research for clinical applications under appropriate regulation and legislation with the hope of alleviating human suffering.

Keeping faith in stem cell research

The Guardian

Tuesday March 25 2008

Cardinal Cormac Murphy-O'Connor (We are made for more, March 24) writes movingly about love and truth, what it is to be human, and the purpose of existence; but he is wrong to exclude atheists from the beauty of his vision. We too can see deep responsibility for others as part of our freedom, just as we can see existence as having a great and wonderful purpose.

However, we do not believe in things because of tradition and dogma. The archbishop writes about meeting a nun who cares for HIV/Aids patients in Zimbabwe, but the Catholic church could drastically cut the incidence of HIV across the world if only it would encourage the use of condoms.

Catholics believe a soul enters the fertilised cell at the moment of conception, thereby making a full human being of it - they believe this despite there being no evidence for it and many philosophical arguments against this view. From this absurd superstition they proscribe much that is beneficial. His Scottish colleague calls the use of hybrid embryos Frankenstein science; a better analogy is with the Frankenstein film where Boris Karloff's monster kills something beautiful and innocent, the little girl, because he does not understand it and fears it. The church is the monster; the beautiful innocent is a science that hurts no one but will save innumerable lives.
Joe Morison
London

What Cardinal Cormac Murphy-O'Connor means by a "free vote" on the human embryology bill - one that is not subject to the Labour whip - is not a free vote at all. At least in so far as Catholic MPs are concerned, it is simply a vote subject to a different whip, namely a Vatican one.

The sight of MPs who are members of a religious sect being urged on by their leader to blackmail the government (by threatening to resign) is one that should raise all sorts of alarm bells with voters. They will be asking who runs Britain - Westminster or Rome? Is Ruth Kelly the MP for Bolton West or Vatican East?
Alistair McBay
National Secular Society

You are right to draw attention to the plight of patients with incurable diseases (Therapeutic cloning offers hope of treatment for Parkinson's; Johnson tries to defuse embryos bill crisis, March 24). However, it is mistaken to believe that embryo stem cell research is the way forward to find cures for these diseases. In spite of the huge sums of money the government has spent so far in supporting this line of research, to date it has yielded nothing of significant therapeutic value.

On the other hand, adult stem cell research is yielding promising results along several lines of research into serious diseases. A clear example is that of bone marrow transplants, and bone marrow harvesting in cancer, a successful application of stem cells in treatment.

Several religious faiths in this country have very grave concerns about the proposed direction of research in the current human fertilisation and embryology bill, and have expressed horror at the idea of mixing animal and human gametes in order to find cures.

If the government professes that we have a democratic multicultural and multi-faith society in this country, it needs to justify its vast expenditure and its continued promotion of a scientific route that is fraught with moral and ethical concerns.
Dr Matthew Thalanany
Colchester, Essex

The Cardinal Archbishop of Glasgow asserts that the fertilisation of animal eggs by human DNA involves the making of babies which are then raided for their constituent parts, demonstrating a lack of respect for human life.

If blastulae are babies then nature itself (or God) demonstrates just such lack of respect since the majority of naturally fertilised human eggs fail to implant and are flushed, unnoticed, into the drains in their thousands every day.
David McBrien
Maidenhead, Berkshire

Surely the logical moral corollary of the premise that there is no animal-human divide is for all who support it to become vegetarian?
Rose Frain
Edinburgh

Story Map of Stem Cells

1. Story map

what does research say about embryonic stem cells, five years after their discovery?whyfiles.org/189stem_cell/

2. Researchers first to map gene that regulates adult stem cell growth

Researchers first to map gene that regulates adult stem cell growth. A new discovery in stem cell research may mean big things for cancer patients in the ...
www.physorg.com/news88013088.html

3. Stem cell plans include creation of embryo bank | The San Diego ...

Oct 5, 2006 ... California's stem cell institute yesterday unveiled its plans for ... insiders met regularly to develop the road map that will guide the ...
www.signonsandiego.com/uniontrib/20061005/news_1n5stem.html -

4. The Stem Cell Story - JDRF Kids Online

The Stem Cell Story. Stem cell research is a topic on which the Juvenile Diabetes Research Foundation International (JDRF) has taken the lead, because it's ...
kids.jdrf.org/index.cfm?fuseaction=home.viewpage&page_id=938B773C-5004-D739-A5CFDCC492B34

Talk about Stem Cells (Video Show)

The Politics of Stem Cell Research
Cell Research...A fertility clinic asks President Bush for advice on Stem Cell Research....jerry zucker stem cell research president bush veto national banana
http://www.youtube.com/watch?v=_QiO6cl8WOk

Autistic Patient After Adult Stem Cell Treatment
Cell Treatment...A young girl with autism shows her amazing improvement following treatment with adult
http://crackle.com/c/Family_Friendly/Autistic_Patient_After_Adult_Ste...

Sam Harris - Stem Cells and Morality
using the case of stem cells. Check out The Science Network at http://thesciencenetwork.org/...Sam Harris Stem Cells science morality atheism
http://www.youtube.com/watch?v=kUwnMX8ht3U

The Charlie Rose Science Series: The Latest on Stem Cell Research
Harvard Stem Cell Institute, George Daley of Children's Hospital Boston and the Harvard Stem Cell Institute, Larry Goldstein, director of the UC San Diego Stem Cell program
http://video.google.com/videoplay?docid=1361375465524090858

Stem Cell Breakthrough
breakthrough in stem cell science, with new technology that lets doctors recover and bank cells from the placenta, too. “By collecting and banking stem cells from
http://www.gofish.com/player.gfp?gfid=30-1154699

Adult Stem Cells (part 6)
6)...An introduction to Adult Stem Cells: The benefits Stem Cell technology can bring to everyday life.
http://www.metacafe.com/watch/734303/adult_stem_cells_part_6/

Stem Cell Primer
Learn all about stem cells in 2-1/2 minutes or less. This is the ending of a weekly show....stem cells embryonic berashis
http://www.youtube.com/watch?v=XoggT8quHYs

Stem Cell Breakthrough
in stem cell science, with new technology that lets doctors recover and bank cells from the placenta, too. �by collecting and banking stem cells from
http://sharkle.com/video/138811/

Adult Stem Cells (part 6)
6)...An introduction to Adult Stem Cells: The benefits Stem Cell technology can bring to
http://tw.video.yahoo.com/video/play?vid=809645

The Stem Cell Debate

THE ISSUES
Access may complicate stem cell study
Elizabeth Cohen: Ethics of stem cell research
Dana Bash: Politics, science, morality
TIME.com: The stem cell debate
TIME.com: When politics and science collide
THE SCIENCE
Blood cells made from stem cells
Stem cells show promise in treating neurological diseases
Stem cells help heal paralyzed rats
THE POLITICS
Bush: Decision made with 'great care'
Reaction mixed to Bush decision
Scientists, senators testify
Foes decry embryo 'slaughter'
Reeve: Fund embryonic stem cell research
House rejects measure allowing human cloning for research
ANALYSIS
Missing the mark on stem cells
Stem cells and a new brain drain
Looking for middle ground in a minefield
Falling behind in the stem cell race?
Embryonic ethics

Everything you need to know for stem cell research (video)

Video Data on Stem Cell Research

1. Every thing you need to know about stem cell....

2. Exploring the Science, Policyand Promise of Stem Cells

3. Human dignity and stem cell research in the EU

3. Stem cells and cancer

4.Stem Cell Biology and...

5. Human Embryonic Stem Cells

Controversy surrounding human embryonic stem cell research

Main article: Stem cell controversy
There exists a widespread controversy over human embryonic stem cell research that emanates from the techniques used in the creation and usage of stem cells. Human embryonic stem cell research is controversial because, with the present state of technology, starting a stem cell line requires the destruction of a human embryo and/or therapeutic cloning. However, recently, it has been shown in principle that embryonic stem cell lines can be generated using a single-cell biopsy similar to that used in preimplantation genetic diagnosis that may allow stem cell creation without embryonic destruction.[29] It is not the entire field of stem cell research, but the specific field of human embryonic stem cell research that is at the centre of an ethical debate.
Opponents of the research argue that embryonic stem cell technologies are a slippery slope to reproductive cloning and can fundamentally devalue human life. Those in the pro-life movement argue that a human embryo is a human life and is therefore entitled to protection.
Contrarily, supporters of embryonic stem cell research argue that such research should be pursued because the resultant treatments could have significant medical potential. It is also noted that excess embryos created for in vitro fertilisation could be donated with consent and used for the research.
The ensuing debate has prompted authorities around the world to seek regulatory frameworks and highlighted the fact that stem cell research represents a social and ethical challenge.

Dolly-10 years later

By Susan Hawes, Writer and Stem Cell Scientist, Monash University

Well, hello, Dolly!
Ten years ago, on February 27, 1997, the scientific journal Nature, published Ian Wilmut and Keith Campbell’s paper announcing the birth of a sheep known as Dolly. Dolly’s genetic parent was not the Scottish Blackface ewe that bore it, but a cell taken from the mammary gland of a Finn Dorset ewe.
Using a technique called nuclear transfer, technicians in the lab, Bill Ritchie and Karen Mycock, laboriously removed the DNA from 430 ewes’ eggs, and inserted into 277 of these the genetic material from mammary epithelial cells. After tricking the reconstructed eggs to divide, and nurturing them in culture medium, 29 became blastocysts, the embryonic stage at which they could be transferred into a surrogate mother. And from one of these came Dolly. This was revolutionary because it showed in the mammal that DNA from an adult cell could once again behave like the DNA of an embryo.
It also showed what English physician William Harvey prophesized in 1651, that “all that is alive comes from the egg.” The egg has a major role in directing how the embryo develops, a characteristic that is exploited to drive development following nuclear transfer.
Dolly, named after Dolly Parton’s famous mammaries, was the catalyst for heated public debates world-wide on the benefits and pitfalls of nuclear transfer, still reverberating today. As BBC correspondent, Pallab Ghosh, reported when Dolly’s birth was announced, “ her ultimate legacy is the start of a scientific revolution.” It is the scientific ramifications from Dolly’s birth and life that are truly profound, and as yet have not fully been realised. Yet, the idea behind these nuclear transfer experiments was not to make copies of existing people as portrayed in movies, nor was it solely to generate patient-specific human embryonic stem cell lines, one touted therapeutic application.
These experiments were undertaken to investigate a basic biological conundrum: during the process of ‘differentiation’, the making of an adult cell, is genetic information lost? That is, are adult cells incapable of being turned back to a more embryonic form? Is the process of differentiation irreversible?
The History The birth of Dolly was really a culmination of a lot of scientific work. As Isaac Newton rightly reiterated: “If I have seen a little further, it is by standing on the shoulders of giants.” It goes back to the late 1800s when Weismann, Roux and Driesch, wanted to understand the process of differentiation. Differentiation is when an embryonic cell becomes many adult cell types. One could visualize the embryonic cell being a bit like clay; it can be molded from one form to another. In contrast, fully differentiated adult cells such as nerve, bone or blood cells, have lost the ability to do many things and have only one primary job to do.
In 1928, Hans Spemann addressed these questions experimentally using salamander eggs. He showed that the nucleus (the part of the cell where our DNA is located) from a very early embryonic cell could within the egg instruct the growth and development of an organism, providing the forerunner experiment for nuclear transfer experiments today.
By 1952, Robert Briggs and Thomas King had transferred the nuclei of an embryo into northern leopard frog eggs and generated tadpoles. They noted that it didn’t work when the nucleus came from older, more ‘adult’ cells, suggesting an older genome might very well be stuck. However, John Gurdon, generated a fully mature African Xenopus frog when he injected non-embryonic nuclei into eggs that had had their own nuclei removed. This work, published in Nature in 1958, was the first to suggest that an adult genome could go backwards in its developmental journey and behave like an embryonic genome again.
While in the public arena it raised the specter of maverick scientists vigorously applying nuclear transfer to make human clones, the research continued. In 1984, James McGrath and Davor Solter developed methods for nuclear transfer in the most common of laboratory animals, the mouse. However, they were only successful transferring nuclei from early embryonic cells. It wasn’t until the birth of Dolly in 1997 that the idea of successful nuclear transfer using an adult mammalian genome was realized.
Where Are We Now? Since Dolly, the use of nuclear transfer of an adult genome has come a long way. It has been successfully applied to many different animal species, including cows, horses, cats, and dogs. Even, noble attempts to save endangered species from extinction by nuclear transfer have been tried, and in some cases resulted in live animals (wild cats, gaur and mouflon sheep). The farming and pharmaceutical industry touts nuclear transfer as a way to make animals with desirable characteristics, such as production of cattle that produce human proteins in their milk. While these feats are considerable, the overall success rate of this procedure remains poor. Indeed, it is remarkable when some nuclear transfer animals, like Dolly, grow up and even have their own offspring- they remain the exception.
Sadly, the legacy for most animals born from a nuclear transfer embryo is severe disabilities and a very short lifespan. So, can nuclear transfer work for human cells? And why would we attempt this? Dolly’s legacy includes the possibility of making patient-specific human embryonic stem cell lines. This would involve taking the DNA from an easily reached cell, such as a skin cell, from a person afflicted with a disease or ailment, and transferring this into an egg which has had its own DNA removed. If the egg can be coaxed into developing into a blastocyst, from this, human embryonic stem cells could be made that have the same DNA as the patient.
Human embryonic stem cells have the potential to generate any adult cell type. If adult cells made from embryonic stem cells, or the embryonic cells themselves, could be used for therapies, the next hurdle will be overcoming their rejection. This problem, of course, is no different than that experienced by recipients of donor organs, who take drugs to muffle their body’s immune system. However, if the human embryonic stem cells are made from nuclear transfer embryos containing a patient’s DNA, then rejection wouldn’t be a problem.
Nowadays, scientists, including Ian Wilmut, argue that this technique is more applicable to making novel human diseased cell lines, a revolutionary way of helping us to understand how diseases develop. This understanding would better enable us to combat disease progression, possibly even to develop novel therapies. Martin Pera, from the Keck School of Medicine, envisages that this “will offer a straightforward route to development of banks of embryonic stem cell lines of a desired phenotype.”
Sadly, the only report of the successful generation of patient-specific cell lines by Woo-Suk Hwang was subsequently exposed as a fraud. No one has yet really shown that a human embryo can develop far enough after nuclear transfer of an adult nuclei to generate embryonic stem cells and the jury is still out as to whether it will. Monkeys, our close relatives, have been born following this process, but only using embryonic and not adult cells for donor nuclei. Furthermore, the procedure was wildly inefficient, highlighting one of the biggest hurdles to human nuclear transfer, the very real problem of acquiring human eggs. In addition to the ethical issues surrounding egg donation, the inefficiency of the nuclear transfer process demands a very high number of eggs providing serious practical constraints.
With the development of this technique relying on scarce human eggs, researchers have suggested using non-human animal eggs to develop cell lines for scientific inquiry; cells that would not be transplanted into a patient, rather used to study disease. Interestingly, this proposition has met with strong objections perhaps more emotional than measured. In Australia, for example, the use of non-human animal eggs for nuclear transfer of human DNA was recently banned.
Gazing into a crystal ball to speculate where these scientific breakthroughs will lead us, we may witness the altering of an adult cell fate being engineered without human eggs. Although this science is in its infancy, if it works, it will be a more practical way of producing patient specific embryonic stem cells. The fruition of this, no matter how long it takes, will be another of Dolly’s remarkable legacies. Nuclear transfer that produced Dolly, is a technique that has allowed us to probe fundamental biological questions and is still critical for us to discover more of the eggs secrets; how they achieve the remarkable feat of guiding embryogenesis, and how they can subvert the fate of a stubborn adult genome.
For these reasons, Dolly was one amazing sheep! Anthropologist Sarah Franklin, attests to this in her reminiscing of meeting with Dolly: “I think what was noticeable about her was that she was such an individual -- somewhat ironically in contrast with her iconic status as a clone. She was very forceful and direct the way 'head ewes' often are, but also, if I can say it without sounding anthropomorphic, a bit coy. Hence, for example, she certainly knew the relationship between cameras and food and not just any food either. Her first foray was to your pockets, the way a dog might do. If she did not get what she wanted she would turn her head away, or even walk to the other end of the paddock as soon as she saw your camera. All the while she would be sneaking a few glances back to see if she was working her wiles on you”. It is interesting that out of all of the animal kingdom, it was the sheep, often parodied as blind followers, rather than leaders, that pioneered this scientific revolution.


Nuclear transfer is a technique in which the nucleus of a somatic cell (any cell of the body apart from the sperm or egg) is transferred into an egg that has had its original nucleus removed. The egg now has the same DNA, or genetic material, as the donor somatic cell. Given the right signals, the egg can be coaxed into developing as if it had been fertilized. The egg would divide to form two cells, then four cells, then eight cells and so on until the blastocyst is formed. Embryonic stem cells can be derived from this blastocyst to create cell lines that are genetically identical to the donor somatic cell. Nuclear transfer may also be referred to as Somatic Cell Nuclear Transfer (SCNT) in other literature.

Clonogenic Multiple Myeloma Progenitors, Stem Cell Properties, and Drug Resistance

Cancer Res. 2008 Jan 1;68(1):190-7.
Matsui W et al.
The Sidney Kimmel Comprehensive Cancer Center and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21231, USA. matsuwi@jhmi.edu
Many agents are active in multiple myeloma, but the majority of patients relapse. This clinical pattern suggests most cancer cells are eliminated, but cells with the clonogenic potential to mediate tumor regrowth are relatively chemoresistant. Our previous data suggested that CD138(+) multiple myeloma plasma cells cannot undergo long-term proliferation but rather arise from clonogenic CD138(neg) B cells. We compared the relative sensitivity of these distinct cell types to clinical antimyeloma agents and found that dexamethasone, lenadilomide, bortezomib, and 4-hydroxycyclophosphamide inhibited CD138(+) multiple myeloma plasma cells but had little effect on CD138(neg) precursors in vitro. We further characterized clonogenic multiple myeloma cells and stained cell lines using the Hoechst side population and Aldefluor assays. Each assay identified CD138(neg) cells suggesting that they possess high drug efflux capacity and intracellular drug detoxification activity. We also found that multiple myeloma cells expressing the memory B-cell markers CD20 and CD27 could give rise to clonogenic multiple myeloma growth in vitro and engraft immunodeficient nonobese diabetes/severe combined immunodeficient mice during both primary and secondary transplantation. Furthermore, both the side population and Aldefluor assays were capable of identifying circulating clonotypic memory B-cell populations within the peripheral blood of multiple myeloma patients. Our results suggest that circulating clonotypic B-cell populations represent multiple myeloma stem cells, and the relative drug resistance of these cells is mediated by processes that protect normal stem cells from toxic injury.

Is Breast Cancer a Stem Cell Disease?

Breast cancer, a stem cell disease.---from curr stem cell res ther.2008.
Breast cancer is a first magnitude problem of public health worldwide. There is increasing evidence that this cancer is originated in and maintained by a small population of undifferentiated cells with self-renewal properties. This small population generates a more differentiated pool of cells which represents the main mass of the tumor, resembling the hierarchical tissue organization of the normal breast. These cancer stem cells seem to share a similar phenotype with their normal counterparts but they display dysfunctional patterns of proliferation and differentiation, and they no longer respond to normal physiological controls that ensure a balanced cellular turnover. The origin of these cancer stem cells is controversial; it is not well known if they are originated from normal stem cells or from more differentiated progenitors where a de novo stem cell program is activated by the oncogenic insult. Here we review the origin of breast cancer stem cells and their role in the pathogenesis of cancer development, together with their implications in breast cancer progression, treatment and prognosis.

Stem cells for myocardial regeneration--a hopeful therapy for heart attacks?

Cardiac myocyte progenitors from adult hearts for myocardial regenerative therapy.
Yang CF et al.

Department of Pediatrics, Taipei Veterans General Hospital, Taipei, Taiwan, R.O.C.
Background: The heart is a highly vascular organ and prolonged interruption of myocardial blood flow initiates events that culminate in cardiac myocyte death. Proposed experimental reparative strategies include harvesting potent cells followed by direct injection into ischemic myocardium to achieve myogenesis and angiogenesis. Methods: Accordingly, we set out to isolate and expand a purified population of adult rat putative cardiomyocyte precursors, and to identify their characteristics in vitro. By using an acute myocardial infarction model and direct cell implantation, we further tested the hypothesis that these cells are an ideal cell source for myocardial regeneration and can enhance cardiac repair after implantation into the ischemic rat heart. Results: We describe here the identification of a subpopulation of primitive cells from rat heart, processing stem cell marker, c-kit and myogenic transcriptional factors, GATA-4 and MEF 2C, and cardiac specific proteins, troponin-I, alpha-sarcomeric actinin and connexin-43. They exhibited a high in vitro proliferative potential. These findings strongly suggest that these cells are putative cardiomyocyte precursors. After transplantation, they were able to be retained and proliferate (13.63 +/- 5.97% after 2 weeks) within the ischemic heart. Progeny of implanted cells migrated along the infarcted scar, reconstituted regenerated cardiomyocytes with incorporation into host myocardium, and inhibited cardiac remodeling with decreased scar formation. Conclusion: Our findings suggest that putative cardiomyocyte precursors isolated from adult heart could potentially be an autologous cell source for myocardial regeneration cell therapy.

Human Skin Cells Reprogrammed Into Embryonic Stem Cells

Science News
Human Skin Cells Reprogrammed Into Embryonic Stem Cells
ScienceDaily (Feb. 12, 2008) — UCLA stem cell scientists have reprogrammed human skin cells into cells with the same unlimited properties as embryonic stem cells without using embryos or eggs.
Led by scientists Kathrin Plath and William Lowry, UCLA researchers used genetic alteration to turn back the clock on human skin cells and create cells that are nearly identical to human embryonic stem cells, which have the ability to become every cell type found in the human body. Four regulator genes were used to create the cells, called induced pluripotent stem cells or iPS cells.
The implications for disease treatment could be significant. Reprogramming adult stem cells into embryonic stem cells could generate a potentially limitless source of immune-compatible cells for tissue engineering and transplantation medicine. A patient's skin cells, for example, could be reprogrammed into embryonic stem cells. Those embryonic stem cells could then be prodded into becoming various cells types -- beta islet cells to treat diabetes, hematopoetic cells to create a new blood supply for a leukemia patient, motor neuron cells to treat Parkinson's disease.
"Our reprogrammed human skin cells were virtually indistinguishable from human embryonic stem cells," said Plath, an assistant professor of biological chemistry, a researcher with the Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research and lead author of the study. "Our findings are an important step towards manipulating differentiated human cells to generate an unlimited supply of patient specific pluripotent stem cells. We are very excited about the potential implications."
The UCLA study confirms the work first reported in late November of researcher Shinya Yamanaka at Kyoto University and James Thompson at the University of Wisconsin. The UCLA research appears Feb. 11, 2008, in an early online edition of the journal Proceedings of the National Academy of the Sciences. The UCLA work was completed at about the same time the Yamanaka and Thomson reports were published. Taken together, the studies demonstrate that human iPS cells can be easily created by different laboratories and are likely to mark a milestone in stem cell-based regenerative medicine, Plath said.
These new techniques to develop stem cells could potentially replace a controversial method used to reprogram cells, somatic cell nuclear transfer (SCNT), sometimes referred to as therapeutic cloning. To date, therapeutic cloning has not been successful in humans. However, top stem cell scientists worldwide stress that further research comparing these reprogrammed cells with stem cells derived from embryos, considered the gold standard, is necessary. Additionally, many technical problems, such as the use of viruses to deliver the four genes for reprogramming, need to be overcome to produce safe iPS cells that can be used in the clinic.
"Reprogramming normal human cells into cells with identical properties to those in embryonic stem cells without SCNT may have important therapeutic ramifications and provide us with another valuable method to develop human stem cell lines," said Lowry, an assistant professor of molecular, cell and developmental biology, a Broad Stem Cell Center researcher and first author of the study. "It is important to remember that our research does not eliminate the need for embryo-based human embryonic stem cell research, but rather provides another avenue of worthwhile investigation."
The combination of four genes used to reprogram the skin cells regulate expression of downstream genes and either activate or silence their expression. The reprogrammed cells were not just functionally identical to embryonic stem cells. They also had identical biological structure, expressed the same genes and could be coaxed into giving rise to the same cell types as human embryonic stem cells.
The UCLA research team included four young scientists recruited to UCLA's new stem cell center in the wake of the passage of Proposition 71 in 2004, which created $3 billion in funding for embryonic stem cell research. The scientists were drawn to UCLA in part because of California's stem cell research friendly atmosphere and the funding opportunities created by Proposition 71. In addition to Plath and Lowry, the team included Amander Clarke, an assistant professor of molecular, cell and developmental biology, and April Pyle, an assistant professor of microbiology, immunology and molecular genetics.
The creation of the human iPS cells is an extension of Plath's work on mouse stem cell reprogramming. Plath headed up one of three research teams that were able to successfully reprogram mouse skin cells into mouse embryonic stem cells. That work appeared in the June 2007 issue of the journal Cell Stem Cell.
Adapted from materials provided by University of California - Los Angeles.

Frequently asked questions about stem cells

Frequently Asked Questions (FAQs)--from NIH.GOV
Basic Questions
What are human embryonic stem cells?
What classes of stem cells are there?
Where do stem cells come from?
Why do scientists want to use stem cell lines?
Healthcare Questions
Why are doctors and scientists so excited about human embryonic stem cells?
Have human embryonic stem cells been used successfully to treat any human diseases yet?
What will be the best type of stem cell to use for therapy?
I have Parkinson's Disease. Is there a clinical trial that I can participate in that uses stem cell as therapy?
Where can I donate umbilical cord stem cells?
Research and Policy Questions
Which research is best to pursue?
Why not use adult stem cells instead of using human embryonic stem cells in research?
What are the NIH Guidelines on the utilization of stem cells derived from human fetal tissue (embryonic germ cells)?
May individual states pass laws to permit human embryonic stem cell research?
Where can I find information about patents obtained for stem cells?
Cell Line Availability and the Registry
I am a scientist funded by the NIH. How many cell lines are available to me, and how do I get them?
I'm interested in purchasing more than one cell line from the NIH Stem Cell Registry. What is known about the status of the cell lines and their availability?
Who owns the cells?
When does NIH anticipate that more stem cells lines will become available?
What policies govern use of stem cell lines from WiCell Research Institute?
Funding Questions
Does NIH fund embryonic stem cell research?
Are there any areas of research involving human pluripotent stem cells that are ineligible for NIH funding?
Can a scientist supported by federal funds conduct research with stem cell lines that are not listed on the NIH Human Embryonic Stem Cell Registry?
What if a scientist is conducting research with both federally fundable and non-federally fundable human embryonic stem cells?
Who is responsible for setting the policy to allow federal money to be used for human embryonic stem cell research?
I am a university research administrator. One of our NIH-funded investigators would like to use a cell line that was created after August 9th, 2001, and it is not eligible for research using federal funds. What should I tell the investigator who wants to work with these cells in his laboratory?
I am an investigator who receives NIH funding, and I am planning to derive new human embryonic stem cell lines. Can I conduct the derivations in my laboratory, or do I need to find a non-university funded laboratory to do this work?
Can you explain what accounting principles are necessary to demonstrate that unallowable charges are not being absorbed by NIH funded research, e.g., indirect costs?
May I use data produced from studies of non-approved human embryonic stem cell (hESC) lines under an NIH-supported project?
Can a DNA clone or plasmid or other research resource originally generated with NIH funds be used in the study of non-approved cell lines?
May a common resource area be created that allows scientists working on unapproved lines and other scientists working on approved lines to use some of the same equipment and common resources (pipettes, glassware, etc.)?

Recent Progresses in Stem Cell Multiponent Differentiation

Ungrin MD, Joshi C, Nica A, Bauwens C, Zandstra PW.

Reproducible, Ultra High-Throughput Formation of Multicellular Organization from Single Cell Suspension-Derived Human Embryonic Stem Cell Aggregates.
PLoS ONE. 2008 Feb 13;3(2):e1565.

Dalby MJ, Andar A, Nag A, Affrossman S, Tare R, McFarlane S, Oreffo RO.

Genomic expression of mesenchymal stem cells to altered nanoscale topographies.
J R Soc Interface. 2008 Feb 12; [Epub ahead of print]

Xu Y, Liu Z, Liu L, Zhao C, Xiong F, Zhou C, Li Y, Shan Y, Peng F, Zhang C.

Neurospheres from rat adipose-derived stem cells could be induced into functional Schwann cell-like cells in vitro.
BMC Neurosci. 2008 Feb 12;9(1):21 [Epub ahead of print]

Arnes JB, Collett K, Akslen LA.

Independent prognostic value of the basal-like phenotype of breast cancer and associations with EGFR and candidate stem cell marker BMI-1.
Histopathology. 2008 Feb;52(3):370-80.

Fan H, Zhang C, Li J, Bi L, Qin L, Wu H, Hu Y.

Gelatin Microspheres Containing TGF-beta3 Enhance the Chondrogenesis of Mesenchymal Stem Cells in Modified Pellet Culture.
Biomacromolecules. 2008 Feb 13; [Epub ahead of print]

[Germ track and stem cells in higher plants]
Tsitol Genet. 2007 Sep-Oct;41(5):67-80. Ukrainian.

Müller EJ, Williamson L, Kolly C, Suter MM.

Outside-in signaling through integrins and cadherins: a central mechanism to control epidermal growth and differentiation?
J Invest Dermatol. 2008 Mar;128(3):501-16.

Hurt EM, Kawasaki BT, Klarmann GJ, Thomas SB, Farrar WL.
(+)CD24(-) prostate cells are early cancer progenitor/stem cells that provide a model for patients with poor prognosis.
Br J Cancer. 2008 Feb 12; [Epub ahead of print]

Bruns I, Steidl U, Fischer JC, Czibere A, Kobbe G, Raschke S, Singh R, Fenk R, Rosskopf M, Pechtel S, von Haeseler A, Wernet P, Tenen DG, Haas R, Kronenwett R.

Pegylated G-CSF mobilizes CD34+ cells with different stem and progenitor subsets and distinct functional properties in comparison with unconjugated G-CSF.
Haematologica. 2008 Feb 11; [Epub ahead of print]

Toth ZE, Leker R, Shahar T, Pastorino S, Szalayova I, Asemenew B, Key S, Parmelee A, Mayer B, Nemeth K, Bratincsak A, Mezey E.

The combination of granulocyte colony stimulatory factor and stem cell factor significantly increases the number of bone marrow derived endothelial cells in brains of mice following cerebral ischemia.
Blood. 2008 Feb 11; [Epub ahead of print]

Quinlan MP, Quatela SE, Philips MR, Settleman J.

Activated Kras, but not Hras or Nras, may initiate tumors of endodermal origin via stem cell expansion.
Mol Cell Biol. 2008 Feb 11; [Epub ahead of print]

McMahon LA, Prendergast PJ, Campbell VA.

A comparison of the involvement of p38, ERK1/2, and PI3K in growth factor-induced chondrogenic differentiation of mesenchymal stem cells.
Biochem Biophys Res Commun. 2008 Feb 8; [Epub ahead of print]

Xu J, Liu X, Jiang Y, Chu L, Hao H, Liu Z, Verfaillie C, Zweier J, Gupta K, Liu Z.

MAPK/ERK signaling mediates VEGF-induced bone marrow stem cell differentiation into endothelial cell.
J Cell Mol Med. 2008 Feb 4; [Epub ahead of print]

Lu ZF, Zandieh Doulabi B, Wuisman PI, Bank RA, Helder MN.

Influence of collagen type II and nucleus pulposus cells on aggregation and differentiation of adipose tissue-derived stem cells.
J Cell Mol Med. 2008 Feb 8; [Epub ahead of print]

Burkert J, Otto W, Wright N.

Side populations of gastrointestinal cancers are not enriched in stem cells.
J Pathol. 2007 Dec 11; [Epub ahead of print]

Li L, Sharma N, Chippada U, Jiang X, Schloss R, Yarmush ML, Langrana NA.

Functional Modulation of ES-Derived Hepatocyte Lineage Cells via Substrate Compliance Alteration.
Ann Biomed Eng. 2008 Feb 12; [Epub ahead of print]

Duan X, Yang L, Dong S, Xin R, Chen G, Guo L.

Characterization of EGFP-labeled mesenchymal stem cells and redistribution of allogeneic cells after subcutaneous implantation.
Arch Orthop Trauma Surg. 2008 Feb 12; [Epub ahead of print]


Kovacevic D, Rodeo SA.

Biological augmentation of rotator cuff tendon repair.
Clin Orthop Relat Res. 2008 Mar;466(3):622-33. Epub 2008 Feb 10.


Jiang J, Chan YS, Loh YH, Cai J, Tong GQ, Lim CA, Robson P, Zhong S, Ng HH.

A core Klf circuitry regulates self-renewal of embryonic stem cells.
Nat Cell Biol. 2008 Feb 10; [Epub ahead of print]