Abstract
Thanks to the Yamanaka transcription factors, pluripotency is recovered in the cell culture dish during in vitro cell reprogramming. Induced pluripotent stem cells (iPSCs) have been introduced to the scientific world as a source of disease models, which are predominantly used in drug discovery and monitoring disease pathophysiology, and as a source of master cell lines for developing cellular therapies. Successfully attaining iPSCs requires careful optimization of many factors, including selection of the transfection method and the appropriate transfection agents; culturing conditions before and after transfection; recovery conditions after transfection, freezing, and thawing; storage conditions; and the choice of cost-effective cell and colony characterization and molecular verification steps. In our optimized procedure, we describe the isolation of peripheral blood mononuclear cells (PBMCs) after blood harvest and their efficient reprogramming into iPSCs using automated electroporator, with piggyBac.
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