TY - JOUR
T1 - Standardization of the Teratoma Assay for Analysis of Pluripotency of Human ES Cells and Biosafety of Their Differentiated Progeny
AU - Gropp, Michal
AU - Shilo, Vitali
AU - Vainer, Gilad
AU - Gov, Miri
AU - Gil, Yaniv
AU - Khaner, Hanita
AU - Matzrafi, Limor
AU - Idelson, Maria
AU - Kopolovic, Juri
AU - Zak, Naomi B.
AU - Reubinoff, Benjamin E.
N1 - Funding Information:
BR is the CSO and holds shares in CellCure Neurosciences Ltd. VS, M. Gov, LM and NBZ were employees of CellCure Neurosciences Ltd. when the study was performed. M. Gropp was funded for this study by CellCure Neurosciences Ltd. This does not alter the authors’ adherence to all the PLOS ONE policies on sharing data and materials.
PY - 2012/9/25
Y1 - 2012/9/25
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84866678032&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0045532
DO - 10.1371/journal.pone.0045532
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C2 - 23049812
AN - SCOPUS:84866678032
VL - 7
JO - PLoS ONE
JF - PLoS ONE
SN - 1932-6203
IS - 9
M1 - e45532
ER -