Standardization of the Teratoma Assay for Analysis of Pluripotency of Human ES Cells and Biosafety of Their Differentiated Progeny |
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| Authors: | Michal Gropp Vitali Shilo Gilad Vainer Miri Gov Yaniv Gil Hanita Khaner Limor Matzrafi Maria Idelson Juri Kopolovic Naomi B Zak Benjamin E Reubinoff |
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| Institution: | 1. The Hadassah Human Embryonic Stem Cell Research Center, The Goldyne Savad Institute of Gene Therapy and Department of Gynecology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.; 2. CellCure NeuroSciences Ltd., Jerusalem, Israel.; 3. The Department of Pathology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.; University of Sao Paulo - USP, Brazil, |
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| Abstract: | Teratoma tumor formation is an essential criterion in determining the pluripotency of human pluripotent stem cells. However, currently there is no consistent protocol for assessment of teratoma forming ability. Here we present detailed characterization of a teratoma assay that is based on subcutaneous co-transplantation of defined numbers of undifferentiated human embryonic stem cells (hESCs) with mitotically inactivated feeder cells and Matrigel into immunodeficient mice. The assay was highly reproducible and 100% efficient when 100,000 hESCs were transplanted. It was sensitive, promoting teratoma formation after transplantation of 100 hESCs, though larger numbers of animals and longer follow-up were required. The assay could detect residual teratoma forming cells within differentiated hESC populations however its sensitivity was decreased in the presence of differentiated cells. Our data lay the foundation, for standardization of a teratoma assay for pluripotency analysis. The assay can also be used for bio-safety analysis of pluripotent stem cell-derived differentiated progeny. |
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