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  • Characterization of embryonic stem-like cells derived from HEK293T cells through miR302/367 expression and their potentiality to differentiate into germ-like cells.

Characterization of embryonic stem-like cells derived from HEK293T cells through miR302/367 expression and their potentiality to differentiate into germ-like cells.

Cytotechnology (2013-10-05)
Long Wang, Haijing Zhu, Jiang Wu, Na Li, Jinlian Hua
ABSTRACT

Human induced pluripotent stem (iPS) cells have great value for regenerative medicine, but are facing problems of low efficiency. MicroRNAs are a recently discovered class of 19-25 nt small RNAs that negatively target mRNAs. miR302/367 cluster has been demonstrated to reprogram mouse and human somatic cells to iPS cells without exogenous transcription factors, however, the repetition and differentiation potentiality of miR302/367-induced pluripotent stem (mirPS) cells need to be improved. Here, we showed overexpression of miR302/367 cluster reprogrammed human embryonic kidney 293T cells into mirPS cells in serum-free N2B27-based medium. The mirPS cells had similar morphology with embryonic stem cells, and expressed pluripotent markers including Oct4, Sox2, Klf4, and Nanog. In addition, through formation of embryoid bodies, various cells and tissues from three germ layers could be determined. Moreover, we examined the potential of mirPS cells differentiating into germ cells both in vitro and in vivo. Taken together, these data might provide a new source of cells and technique for the investigation of the mechanisms underlying reprogramming and pluripotency.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Retinoic acid, ≥98% (HPLC), powder
Sigma-Aldrich
Anti-SOX-2 Antibody, clone 6F1.2, Chemicon®, from mouse
Sigma-Aldrich
Anti-c-myc Antibody, Chemicon®, from chicken
Sigma-Aldrich
Anti-Glial Fibrillary Acidic Protein (GFAP) Antibody, serum, Chemicon®
Sigma-Aldrich
Anti-KLF4 Antibody, Chemicon®, from rabbit