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Abstract
The International Stem Cell Initiative compared several commonly used approaches to assess human pluripotent stem cells (PSC). PluriTest predicts pluripotency through bioinformatic analysis of the transcriptomes of undifferentiated cells, whereas, embryoid body (EB) formation in vitro and teratoma formation in vivo provide direct tests of differentiation. Here we report that EB assays, analyzed after differentiation under neutral conditions and under conditions promoting differentiation to ectoderm, mesoderm, or endoderm lineages, are sufficient to assess the differentiation potential of PSCs. However, teratoma analysis by histologic examination and by TeratoScore, which estimates differential gene expression in each tumor, not only measures differentiation but also allows insight into a PSC’s malignant potential. Each of the assays can be used to predict pluripotent differentiation potential but, at this stage of assay development, only the teratoma assay provides an assessment of pluripotency and malignant potential, which are both relevant to the pre-clinical safety assessment of PSCs.
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1 Centre for Stem Cell Biology, Department of Biomedical Science, University of Sheffield, Western Bank, Sheffield, UK; UCLA School of Medicine, Department of Biological Chemistry, Los Angeles, CA, USA
2 Centre for Stem Cell Biology, Department of Biomedical Science, University of Sheffield, Western Bank, Sheffield, UK
3 The Azrieli Center for Stem Cells and Genetic Research, Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University, Jerusalem, Israel
4 CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria; Department of Laboratory Medicine, Medical University of Vienna, Vienna, Austria; Max Planck Institute for Informatics, Saarland Informatics Campus, Saarbrücken, Germany
5 WiCell Research Institute Inc., Madison, WI, USA
6 Institute of Reconstructive Neurobiology, LIFE and BRAIN Center, University of Bonn Medical Faculty, Bonn, Germany
7 Department of Pathology, The University of Kansas School of Medicine, Kansas City, KS, USA
8 Murdoch Childrens Research Institute, Royal Children’s Hospital, Parkville, VIC, Australia; Department of Anatomy and Developmental Biology, Monash University, Clayton, Australia
9 CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria
10 UK Stem Cell Bank, Advanced Therapies Division, NIBSC-MHRA, Blanche Ln, South Mimms, Potters Bar, UK; The Francis Crick Institute, London, UK
11 Genome Institute of Singapore, Singapore, Singapore
12 Jackson Laboratory, Bar Harbor, ME, USA; Siriraj Center of Excellence in Stem Cell Research, Mahidol University, Bangkok, Thailand
13 The Scripps Research Institute, La Jolla, CA, USA
14 Department of Genome Regulation, Max-Planck-Institute for Molecular Genetics, Berlin, Germany; Zentrum für Integrative Psychiatrie, Universitätsklinikum Schleswig-Holstein, Kiel, Germany
15 Leiden University Medical Center, Leiden, The Netherlands
16 Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto, Japan
17 Bioprocessing Technology Institute, Singapore, Singapore
18 UK Stem Cell Bank, Advanced Therapies Division, NIBSC-MHRA, Blanche Ln, South Mimms, Potters Bar, UK
19 Stem Cells Australia, The University of Melbourne, Parkville, VIC, Australia; Department of Anatomy and Neurosciences, The University of Melbourne, Parkville, VIC, Australia; Florey Neuroscience and Mental Health Institute, Parkville, VIC, Australia; Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia; The Jackson Laboratory, Bar Harbor, ME, USA
20 Department of Obstetrics and Gynecology, and the Hadassah Human Embryonic Stem Cells Research Center, The Goldyne-Savad Institute of Gene Therapy, Hadassah-Hebrew University Hospital, Jerusalem, Israel
21 Genome Institute of Singapore, Singapore, Singapore; The Jackson Laboratory for Genomic Medicine, Farmington, CT, USA; Department of Genetics and Genome Sciences, University of Connecticut Health Center, Farmington, CT, USA
22 Program in Developmental and Stem Cell Biology, Hospital for Sick Children, Toronto, Canada; Department of Molecular Genetics, University of Toronto, Toronto, Canada
23 Zentrum für Integrative Psychiatrie, Universitätsklinikum Schleswig-Holstein, Kiel, Germany
24 Siriraj Center of Excellence in Stem Cell Research, Mahidol University, Bangkok, Thailand; Max Planck Society, Munich, Germany
25 UK Stem Cell Bank, Advanced Therapies Division, NIBSC-MHRA, Blanche Ln, South Mimms, Potters Bar, UK; The International Stem Cell Banking Initiative, Barley, Herts, UK
26 Laboratory of Embryonic Stem Cell research, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan
27 Center for iPS cell Research and Application, Kyoto University, Kyoto, Japan
28 Center for iPS cell Research and Application, Kyoto University, Kyoto, Japan; Gladstone Institute of Cardiovascular Disease, San Francisco, CA, USA