Varga Eszter, Hansen Marten, Wüst Tatjana, von Lindern Marieke, van den Akker Emile
Sanquin Research, Dept. Hematopoiesis, Amsterdam, The Netherlands; Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands.
Sanquin Research, Dept. Hematopoiesis, Amsterdam, The Netherlands; Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands.
Stem Cell Res. 2017 Dec;25:30-33. doi: 10.1016/j.scr.2017.10.001. Epub 2017 Oct 12.
Peripheral blood mononuclear cells were isolated and cultured to a pure pro-EBL population and reprogrammed using episomal plasmids. The pluripotency of transgene-free induced pluripotent stem cell (iPSC) line was verified by the expression of pluripotency-associated markers and by in vitro spontaneous differentiation towards the 3 germ layers. The iPSC line showed normal karyotype. Peripheral blood is a non-invasive easy accessible cell source and combined with EBL outgrowth in vitro, a routine process obtaining sufficient amount of homogenous cells can be obtained within a week. Using episomal delivery, pro-EBLs can be reprogrammed in a transgene-free, cost effective system.
分离外周血单个核细胞并培养至纯原幼红细胞白血病(pro-EBL)群体,然后使用游离型质粒进行重编程。通过多能性相关标志物的表达以及向三个胚层的体外自发分化,验证了无转基因诱导多能干细胞(iPSC)系的多能性。该iPSC系显示出正常的核型。外周血是一种非侵入性且易于获取的细胞来源,结合体外原幼红细胞白血病细胞生长,一周内即可获得足够数量的同质细胞的常规流程。使用游离型载体递送,原幼红细胞白血病细胞可在无转基因、成本效益高的系统中进行重编程。