Mou Hongmei, Vinarsky Vladimir, Tata Purushothama Rao, Brazauskas Karissa, Choi Soon H, Crooke Adrianne K, Zhang Bing, Solomon George M, Turner Brett, Bihler Hermann, Harrington Jan, Lapey Allen, Channick Colleen, Keyes Colleen, Freund Adam, Artandi Steven, Mense Martin, Rowe Steven, Engelhardt John F, Hsu Ya-Chieh, Rajagopal Jayaraj
Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Division of Pediatric Pulmonary Medicine, Massachusetts General Hospital for Children, Boston, MA 02114, USA.
Center for Regenerative Medicine, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Massachusetts General Hospital, Boston, MA 02114, USA.
Cell Stem Cell. 2016 Aug 4;19(2):217-231. doi: 10.1016/j.stem.2016.05.012. Epub 2016 Jun 16.
Functional modeling of many adult epithelia is limited by the difficulty in 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 find 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 signaling inhibition in a feeder-free culture system, we have been able to expand airway basal stem cells from multiple species. Expanded cells can produce functional airway epithelium physiologically responsive to clinically relevant drugs, such as CFTR modulators. This approach is effective for the clonal expansion of single human cells and for basal cell populations from epithelial tissues from all three germ layers and therefore may be broadly applicable for modeling of epithelia.
许多成人上皮组织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，我们表明对SMAD信号通路的双重抑制能够使原代上皮基底细胞群体强劲扩增。我们发现，TGFβ/BMP/SMAD通路信号在几种上皮组织的管腔和基底上层细胞中被强烈激活，但在p63+基底细胞中受到抑制。在气道上皮中，SMAD信号促进分化，其抑制导致干细胞增生。在无饲养层培养系统中使用双重SMAD信号抑制，我们已经能够扩增来自多个物种的气道基底干细胞。扩增的细胞可以产生对临床相关药物（如CFTR调节剂）具有生理反应的功能性气道上皮。这种方法对于单个人类细胞的克隆扩增以及来自所有三个胚层的上皮组织的基底细胞群体都是有效的，因此可能广泛适用于上皮组织的建模。
