Lili Du1, Ge Lin, Guangxiu Lu. 1. Institute of Reproductive & Stem Cell Engineering, Central South University, Changsha 410078, China.
Abstract
OBJECTIVE: To generate and identify the induced pluripotent stem cells (iPSCs) from human embryonic fibroblast cells (hEFs) by introducing 4 defined factors. METHODS: We introduced 4 factors (Oct4, Sox2, Nanog,and Lin28) into hEFs by lentivirus infection. The iPSCs generated from this method were analyzed in many aspects, including surface antigens, gene expression, and telomerase activity, differentiation ability in vivo and in vitro and the patterns of short tandem repeat (STR). RESULTS: The human iPSCs generated from this study were positive for alkaline phosphatase (AKP) staining, expressed hESCs-specific surface antigens, and exhibited high telomerase activity. They also possessed the ability to differentiate into 3-germ layers both in vivo and in vitro. STR analysis indicated that the human iPSCs were derived from the donor material. CONCLUSION: The iPSCs lines can be obtained from human embryonic fibroblast by introducing 4 factors. Such human iPSCs should be useful in the study of epigenetic reprogramming and pluripotency maintenance.
OBJECTIVE: To generate and identify the induced pluripotent stem cells (iPSCs) from human embryonic fibroblast cells (hEFs) by introducing 4 defined factors. METHODS: We introduced 4 factors (Oct4, Sox2, Nanog,and Lin28) into hEFs by lentivirus infection. The iPSCs generated from this method were analyzed in many aspects, including surface antigens, gene expression, and telomerase activity, differentiation ability in vivo and in vitro and the patterns of short tandem repeat (STR). RESULTS: The human iPSCs generated from this study were positive for alkaline phosphatase (AKP) staining, expressed hESCs-specific surface antigens, and exhibited high telomerase activity. They also possessed the ability to differentiate into 3-germ layers both in vivo and in vitro. STR analysis indicated that the human iPSCs were derived from the donor material. CONCLUSION: The iPSCs lines can be obtained from human embryonic fibroblast by introducing 4 factors. Such human iPSCs should be useful in the study of epigenetic reprogramming and pluripotency maintenance.
Authors: Papia Chakraborty; F William Buaas; Manju Sharma; Elizabeth Snyder; Dirk G de Rooij; Robert E Braun Journal: Stem Cells Date: 2014-04 Impact factor: 6.277