REPROGRAMMING NORMAL AND CANCEROUS HUMAN CELL LINES INTO HUMAN INDUCED POLURIPOTENT STEM CELLS BY CO-ELECTROPORATION WITH LIVING XENOPUS LAEVIS FROG OOCYTES

摘要

Using electroporation, it is possible to activate the natural reprogramming potential of living Xenopus laevis oocytes and pass it on to donor cells placed with eggs in one electroporation chamber. We demonstrated that co-electroporation at 150 v/cm/25 μF of mature oocytes with ^˜10⁵ cells/ml of suspension of various normal and cancerous human cell lines, such as bone marrow stromal cells, foreskin fibroblasts, pre-adipocytes, CD4+ T-lymphocytes, cheek cells, cervical carcinoma (HeLa) cells and breast adenocarcinoma (MCF-7) cells, reprograms donor cells into iPSc-like cells, which form colonies on irradiated MEF feeders. The iPSc-like cells generated by this study resemble human embryonic stem cells in colony morphology and expression of stem cell-associated transcription factors, including Oct3/4, Nanog, SOX-2, Rex-1, TRA-1-60 and SSEA-1. New method obviates the use of retroviral or lentiviral gene delivery vectors and other “non-parental” reprogramming approaches.