Long-Term In Vitro Expansion of Epithelial Stem Cells Enabled by Pharmacological Inhibition of PAK1-ROCK-Myosin II and TGF-β Signaling.
Long-Term In Vitro Expansion of Epithelial Stem Cells Enabled by Pharmacological Inhibition of PAK1-ROCK-Myosin II and TGF-β Signaling.
复制标题
DOI：
10.1016/j.celrep.2018.09.072
复制
发表时间：
2018-10-16
期刊：
影响因子：
8.8
通讯作者：
Wang T
中科院分区：
文献类型：
--
作者：
Zhang C;Lee HJ;Shrivastava A;Wang R;McQuiston TJ;Challberg SS;Pollok BA;Wang T
尽管在体内具有很强的自我更新能力，但位于各种组织中的上皮干细胞和祖细胞在体外无饲料条件下会扩增几代，然后就会出现生长停滞。在这里，我们描述了EpiX方法，该方法利用抑制PAK1-ROCK-Myosin II和TGF-β信号传导的小分子，在没有饲养细胞的情况下，实现了来自皮肤、气道、乳腺和前列腺的人上皮干细胞和祖细胞超过一万亿倍的扩增。转录组学和表观基因组学研究表明，这种情况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与其组织起源一致的成熟上皮细胞。全基因组测序显示，在体外广泛扩增后，细胞保持了显著的基因组完整性，而没有获得致瘤性。EpiX技术为开发组织驻留上皮干细胞和祖细胞用于再生医学的潜力提供了一种解决方案。Zhang等人筛选了小分子集合，发现在低钙条件下，TGF-β和PAK1-ROCK-Myosin II的药理抑制支持上皮干细胞在2D格式下的扩展扩张。这种方法增强了组织驻留上皮干细胞用于细胞治疗的潜力。
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. Zhang et al. screen a small-molecule collection and find that pharmacologic inhibition of TGF-β and PAK1-ROCK-Myosin II, in low calcium conditions, supports extended expansion of epithelial stem cells in 2D format. This approach enhances the potential of tissue-resident epithelial stem cells for cell therapy.
登录
查看更多内容
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
14.9
作者：
Gordon K;Clouaire T;Bao XX;Kemp SE;Xenophontos M;de Las Heras JI;Stancheva I
通讯作者：
Stancheva I
DOI：
10.1146/annurev-pathol-121808-102144
发表时间：
2010
期刊：
Annual review of pathology
影响因子：
--
作者：
Coppé JP;Desprez PY;Krtolica A;Campisi J
通讯作者：
Campisi J
DOI：
10.1126/science.1242281
发表时间：
2014-06-13
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Blanpain C;Fuchs E
通讯作者：
Fuchs E
影响因子：
64.5
作者：
Huch M;Gehart H;van Boxtel R;Hamer K;Blokzijl F;Verstegen MM;Ellis E;van Wenum M;Fuchs SA;de Ligt J;van de Wetering M;Sasaki N;Boers SJ;Kemperman H;de Jonge J;Ijzermans JN;Nieuwenhuis EE;Hoekstra R;Strom S;Vries RR;van der Laan LJ;Cuppen E;Clevers H
通讯作者：
Clevers H