Propagenix, 9605 Medical Center Drive, Suite 325, Rockville, MD 20850, USA.
Department of Genetics, Washington University School of Medicine, St. Louis, MO 63108, USA; The Edison Family Center for Genome Sciences and Systems Biology, Washington University School of Medicine, St. Louis, MO 63108, USA.
Cell Rep. 2018 Oct 16;25(3):598-610.e5. doi: 10.1016/j.celrep.2018.09.072.
Despite substantial self-renewal capability in vivo, epithelial stem and progenitor cells located in various tissues expand for a few passages in vitro in feeder-free condition before they succumb to growth arrest. Here, we describe the EpiX method, which utilizes small molecules that inhibit PAK1-ROCK-Myosin II and TGF-β signaling to achieve over one trillion-fold expansion of human epithelial stem and progenitor cells from skin, airway, mammary, and prostate glands in the absence of feeder cells. Transcriptomic and epigenomic studies show that this condition helps epithelial cells to overcome stresses for continuous proliferation. EpiX-expanded basal epithelial cells differentiate into mature epithelial cells consistent with their tissue origins. Whole-genome sequencing reveals that the cells retain remarkable genome integrity after extensive in vitro expansion without acquiring tumorigenicity. EpiX technology provides a solution to exploit the potential of tissue-resident epithelial stem and progenitor cells for regenerative medicine.
尽管体内的上皮干细胞和祖细胞具有很强的自我更新能力，但在无饲养细胞的条件下，它们在体外只能有限地传代扩增几次，随后就会进入生长停滞状态。在这里，我们描述了一种 EpiX 方法，该方法利用小分子抑制 PAK1-ROCK-Myosin II 和 TGF-β 信号通路，从而能够在没有饲养细胞的情况下，将来自皮肤、气道、乳腺和前列腺等组织的人上皮干细胞和祖细胞扩增超过一万亿倍。转录组学和表观基因组学研究表明，这种条件有助于上皮细胞克服持续增殖的压力。EpiX 扩增的基底上皮细胞分化为成熟的上皮细胞，与它们的组织来源一致。全基因组测序显示，在没有获得致瘤性的情况下，经过大量体外扩增，细胞仍然保持着显著的基因组完整性。EpiX 技术为开发组织驻留的上皮干细胞和祖细胞的潜力用于再生医学提供了一种解决方案。
