Efficient generation of transgene-free induced pluripotent stem cells from normal and neoplastic bone marrow and cord blood mononuclear cells
Efficient generation of transgene-free induced pluripotent stem cells from normal and neoplastic bone marrow and cord blood mononuclear cells
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DOI：
10.1182/blood-2010-07-298331
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发表时间：
2011-04-07
期刊：
影响因子：
20.3
通讯作者：
Slukvin, Igor I.
中科院分区：
文献类型：
--
作者：
Hu, Kejin;Yu, Junying;Slukvin, Igor I.
将血细胞重编程为诱导多能干细胞（iPSC）为体外血液疾病建模提供了一种新的工具。然而，当前重编程技术的众所周知的局限性包括低效率、慢动力学、以及转基因整合和残余表达。在本研究中，我们已经证明，可以使用非整合附加型载体从人BM和CB单核细胞产生不含转基因和载体序列的iPSC。与成纤维细胞的重编程相比，本文描述的重编程效率高出100倍，发生时间快1-3周，并且不需要分离祖细胞或多轮转染。血液来源的iPSC系缺乏IGH和TCR的重排，表明它们的来源是非B或非T淋巴细胞。当在OP 9上共培养时，血液来源的iPSC可以分化回血细胞，尽管与成纤维细胞来源的iPSC相比效率较低。我们还从患有慢性髓性白血病（CML）的患者的BM中产生了无转基因的iPSC。CMLiPSC显示出在骨髓样品中鉴定的独特的复杂染色体易位，同时显示出典型的胚胎干细胞表型和多能分化潜能。这种方法提供了一个机会，探索库存的正常和患病的CB和BM样本，而没有与基于病毒的方法相关的限制。（血。2011;117（14）：e109-e119）
Reprogramming blood cells to induced pluripotent stem cells (iPSCs) provides a novel tool for modeling blood diseases in vitro. However, the well-known limitations of current reprogramming technologies include low efficiency, slow kinetics, and transgene integration and residual expression. In the present study, we have demonstrated that iPSCs free of transgene and vector sequences could be generated from human BM and CB mononuclear cells using nonintegrating episomal vectors. The reprogramming described here is up to 100 times more efficient, occurs 1-3 weeks faster compared with the reprogramming of fibroblasts, and does not require isolation of progenitors or multiple rounds of transfection. Blood-derived iPSC lines lacked rearrangements of IGH and TCR, indicating that their origin is non-B- or non-T-lymphoid cells. When cocultured on OP9, blood-derived iPSCs could be differentiated back to the blood cells, albeit with lower efficiency compared to fibroblast-derived iPSCs. We also generated transgene-free iPSCs from the BM of a patient with chronic myeloid leukemia (CML). CMLiPSCs showed a unique complex chromosomal translocation identified in marrow sample while displaying typical embryonic stem cell phenotype and pluripotent differentiation potential. This approach provides an opportunity to explore banked normal and diseased CB and BM samples without the limitations associated with virus-based methods. (Blood. 2011;117(14):e109-e119)