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Broader implications of defining standards for the pluripotency of iPSCs

Broader implications of defining standards for the pluripotency of iPSCs

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Published by Nora Yücel

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Published by: Nora Yücel on Sep 08, 2010
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10/21/2010

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Cell Stem Cell
Letter
Broader Implications of Defining Standardsfor the Pluripotency of iPSCs
George Q. Daley,
1,2,
*M. William Lensch,
Rudolf Jaenisch,
3
 Alex Meissner,
2,4
Kathrin Plath,
5
and Shinya Yamanaka
6,7
1
Children’s Hospital Boston, Division of Hematology/Oncology, 300 Longwood Avenue, Boston, MA 02115, USA 
2
Harvard Stem Cell Institute, 42 Church Street, Cambridge, MA 02138, USA 
3
Whitehead Institute for Biomedical Research and Massachusetts Institute of Technology, Department of Biology, 9 Cambridge Center,Cambridge, MA 02142, USA 
4
Department of Stem Cell and Regenerative Biology, Harvard University, 42 Church Street, Cambridge, MA 02138, USA 
5
University of California, School of Medicine, Department of Biological Chemistry, The Broad Center for Regenerative Medicineand Stem Cell Research, 615 Charles E. Young Drive South, Los Angeles, CA 90095, USA 
6
Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan
7
Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA *Correspondence:george.daley@childrens.harvard.eduDOI 10.1016/j.stem.2009.02.009
We read with great interest the excellentreview byMaherali and Hochedlinger(2008)that recommends standards forcharacterization of pluripotent stem celllines, especially the many new lines beinggenerated using factor-based reprogram-ming techniques (induced pluripotentstem cells, or iPSCs). Of note was thesuggestionthatiPSCsshouldbeassessedfor ‘‘functional differentiation through thehighest-stringency test acceptable.’Formurine iPSCs, this means germline trans-mission following blastocyst chimerism,and for human iPSCs this means assess-ment of teratoma pathology. Given thefastpaceofdiscoveryinthefield,thevalueand relevance of time-consuming charac-terization of cell lines are bound to bedebated. We’d like to highlight what’s atrisk when the pressure for rapid publica-tion erodes the imperative for applyingrigorous and uniform standards beforeassigning the label ‘‘iPSC’’ to novel celllines.The term ‘‘pluripotency’’ can be as-signed according to lax or stringentcriteria. A diverse array of stem cell typeshave been labeled pluripotent: multipo-tential adult progenitor cells (MAPCs);amniotic fluid-derived stem cells (AFS);marrow-isolated adult multilineage induc-ible cells (MIAMI); testes-derived stemcells;andavarietyofembryonicstemcellsderived by parthenogenesis, blastomereculture, and somatic cell nuclear transfer.In the loosest sense, a pluripotent cellincludes in its progeny elements of allthree embryonic germ layers (ectoderm,endoderm, and mesoderm), regardlessof experimental context. In the strictestsense, pluripotency pertains to cellswhose progeny can reconstitute an entireorganism, and is measured most strin-gently in the mouse using tetraploidembryo complementation (a standardachieved by only a limited subset of ESCs). To date, murine iPSCs have notyieldedlive pupsin thetetraploid comple-mentation assay, and thus the standardroutinely applied is transmission of cellsthrough the germline of chimeric animalsto yield live pups (and not just gametes).Given practical and ethical limitations ontesting of human embryonic stem cells,the gold standard for assessing pluripo-tency is the capacity to generate well-differentiated teratomas following injec-tion into immunodeficient mice ( Brivanlouet al., 2003 ). Although nonquantitativeand subjective, teratoma histology in thehands of a skilled pathologist can distin-guish tumors composed predominantlyof poorly differentiated neuroectodermalelements from cystic masses composedof well-differentiated tissue from all threeembryonic germ layers. The formerbehave as malignancies and are akin toteratocarcinomas, while the latter behaveas encapsulated, benign masses and aretrue teratomas that arise from pluripotentstem cells ( Lensch et al., 2007 ).Importantly, in the mouse there areseveraltypesofembryo-derivedstemcellsthat all share the most basic capacity fordifferentiation into all three germ layers:classicalESCs,epiblast-derivedstemcells(EpiSCs), and Fibroblast Growth Factor Activin/Bio cultivated stem cells (FAB-SCs), among them. These different typesof embryo-derived stem cells behavedifferently when subjected to specificin vivo assays, in some instances formingteratomasbutfailingthecriterionofblasto-cystchimerismandgermlinetransmission.Choosing to ignore the need to clearlyestablish the behavior of these and othernovelclassesofstemcellshasbothscien-tific (and we might point out, political)consequences. MAPCs and AFS cellshave been touted as viable alternatives tohuman ESCs. While these cells may bevaluable for generating differentiated celltypes in vitro, they meet very differentcriteria for pluripotency. For nearly adecade after human ESCs were firstisolated, stem cell scientists operatedunder the presumption that human ESCswere equivalent to mouse ESCs. Recentevidence gleaned from a careful compar-ison of growth factor requirements andbehaviorinvariousassaysofdifferentiationnow indicates that human ESCs are mostsimilar to EpiSCs. How much do we knowabout iPSCs given that factor-based re-programmingisbarelymorethantwoyearsold?TheinitialiPSCsisolatedbyTakahashiandYamanaka(2006)behavedquitediffer-ently fromthe subsequently cultured lines;the initial iPSC lines chimerized embryosbut didn’t yield live pups or chimerize thegermline.Partiallyreprogrammedcoloniescan be identified and pushed toward fullpluripotency with subsequent chemicaltreatment Meissner et al., 2008 ), andhuman colonies that might be mistakenfor faithfully reprogrammed iPSCs indeedfail a number of criteria for pluripotency,includingmarkerexpressionandformationof highly differentiated teratomas. In fact,the mere detection of cell-type-specificmarkers on cells grown in culture isa less stringent criterion for functionaldifferentiation than the presence of 
200
Cell Stem Cell
4
, March 6, 2009
ª
2009 Elsevier Inc.

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