Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells.
Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells.
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DOI：
10.1016/j.stem.2016.05.012
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发表时间：
2016-08-04
期刊：
影响因子：
23.9
通讯作者：
Rajagopal J
中科院分区：
文献类型：
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作者：
Mou H;Vinarsky V;Tata PR;Brazauskas K;Choi SH;Crooke AK;Zhang B;Solomon GM;Turner B;Bihler H;Harrington J;Lapey A;Channick C;Keyes C;Freund A;Artandi S;Mense M;Rowe S;Engelhardt JF;Hsu YC;Rajagopal J
许多成体上皮细胞的功能建模受到在培养中维持相关干细胞群体的困难的限制。在这里，我们表明，SMAD信号通路的双重抑制能够使初级上皮基底层细胞种群强劲扩张。我们发现转化生长因子β/骨形态发生蛋白/SmadD信号通路在几种上皮的管腔和基底上细胞中被强烈激活，而在p63+的基底细胞中被抑制。在呼吸道上皮细胞中，SMAD信号促进分化，其抑制导致干细胞增殖。在无饲养层培养系统中使用双重SMAD抑制，我们能够扩增来自多个物种的呼吸道基础干细胞。扩增的细胞可以产生功能性的呼吸道上皮，对临床相关药物如CFTR调节剂有生理反应。这种方法对于人类单个细胞的克隆扩增和来自所有三个生殖层的上皮组织的基底细胞群都是有效的，因此可能广泛适用于上皮细胞的建模。MOU等人的研究成果。结果表明，小分子介导的SMAD信号抑制允许以2D形式长期无饲养层培养不同功能的上皮基础干细胞。这种方法提供了一个简便的患者特定的上皮性疾病建模平台，如从囊性纤维化患者的非侵入性获取的标本中扩增呼吸道上皮所示。
Functional modeling of many adult epithelia is limited by the difficulty of maintaining relevant stem cell populations in culture. Here, we show that dual inhibition of SMAD signaling pathways enables robust expansion of primary epithelial basal cell populations. We found that TGFβ/BMP/SMAD pathway signaling is strongly activated in luminal and suprabasal cells of several epithelia, but suppressed in p63+ basal cells. In airway epithelium, SMAD signaling promotes differentiation, and its inhibition leads to stem cell hyperplasia. Using dual SMAD inhibition in a feeder-free culture system we were able to expand airway basal stem cells from multiple species. Expanded cells can produce functional airway epithelium that is physiologically responsive to clinically relevant drugs such as CFTR modulators. This approach is effective for clonal expansion of single human cells and for basal cell populations from epithelial tissues from all three germ layers, and may therefore be broadly applicable for modeling of epithelia. Mou et al. show that small molecule-mediated SMAD signaling inhibition allows prolonged feeder-free culture of diverse functional epithelial basal stem cells in a 2D format. This methodology provides a facile patient-specific epithelial disease modeling platform, as shown by expanding airway epithelium from non-invasively obtained specimens from cystic fibrosis patients.