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Publication : Generation of mouse-induced pluripotent stem cells with plasmid vectors.

First Author  Okita K Year  2010
Journal  Nat Protoc Volume  5
Issue  3 Pages  418-28
PubMed ID  20203661 Mgi Jnum  J:346830
Mgi Id  MGI:6511365 Doi  10.1038/nprot.2009.231
Citation  Okita K, et al. (2010) Generation of mouse-induced pluripotent stem cells with plasmid vectors. Nat Protoc 5(3):418-28
abstractText  Reprogramming of somatic cells into pluripotent stem cells has been reported by introducing a combination of several transcription factors (Oct3/4, Sox2, Klf4 and c-Myc). The induced pluripotent stem (iPS) cells from patient's somatic cells could be a useful source for drug discovery and cell transplantation therapies. However, to date, most iPS cells were made using viral vectors, such as retroviruses and lentiviruses. Here we describe an alternative method to generate iPS cells from mouse embryonic fibroblasts (MEFs) by continual transfection of plasmid vectors. This protocol takes around 2 months to complete, from MEF isolation to iPS cell establishment. Although the reprogramming efficiency of this protocol is still low, the established iPS cells are most likely free from plasmid integration. This virus-free technique reduces the safety concern for iPS cell generation and application, and provides a source of cells for the investigation of the mechanisms underlying reprogramming and pluripotency.
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