Directed differentiation of human pluripotent stem cells into diverse organ-specific mesenchyme of the digestive and respiratory systems.
Directed differentiation of human pluripotent stem cells into diverse organ-specific mesenchyme of the digestive and respiratory systems.
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DOI：
10.1038/s41596-022-00733-3
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发表时间：
2022-11
期刊：
影响因子：
14.8
通讯作者：
Zorn, Aaron M.
中科院分区：
文献类型：
--
作者：
Kishimoto, Keishi;Iwasawa, Kentaro;Sorel, Alice;Ferran-Heredia, Carlos;Han, Lu;Morimoto, Mitsuru;Wells, James M.;Takebe, Takanori;Zorn, Aaron M.
食道、肺、肝脏和胃等内脏器官的发育是通过胎儿前肠内胚层和相邻中胚层细胞之间的相互信号相互作用来协调的。虽然最近在体外重现发育信号方面取得的成功使人多能干细胞（hPSC）能够分化为各种类型的器官特异性内胚层上皮，但器官特异性间充质的生成受到的关注要少得多。这是持续设计复杂人体组织的主要限制。在这里，我们描述了利用小鼠前肠器官发生的单细胞转录组学阐明的信号网络和分子标记将 hPSC 分化为不同类型的器官特异性中胚层的方案。基于既定方法，用视黄酸 (RA) 或 RA 与 Hedgehog (HH) 激动剂一起处理的 hPSC 衍生侧板中胚层 (LPM) 在培养 4 天后分别生成后部或前部前肠内脏中胚层 (pFG-SpM 或 aFG-SpM)。在培养物中第 4 天至第 7 天，通过组合激活或抑制 WNT、BMP、RA 或 HH 途径，这些物质被引导至器官特异性间充质谱系。到第 7 天，培养物富集了具有不同分子特征的不同类型的中胚层：60-90% 纯肝横隔/类间皮 (STM/mesothelium)、70-80% 纯肝样成纤维细胞 (LF) 和约 35% 呼吸样中胚层 (RM)、胃样中胚层 (GM) 或食管样中胚层 (EM) 群体。该协议可以由具有中等分化 hPSC 经验的任何人执行，并提供一个研究人类中胚层发育的新平台，并可用于设计更复杂的前肠组织，用于疾病建模和再生医学。
Development of visceral organs such as the esophagus, lung, liver and stomach are coordinated by reciprocal signaling interactions between the endoderm and adjacent mesoderm cells in the fetal foregut. While the recent successes in recapitulating developmental signaling in vitro has enabled the differentiation of human pluripotent stem cells (hPSCs) into various types of organ-specific endodermal epithelium, the generation of organ-specific mesenchyme has received much less attention. This is a major limitation in ongoing efforts to engineer complex human tissue. Here, we describe protocols to differentiate hPSCs into different types of organ-specific mesoderm leveraging signaling networks and molecular markers elucidated from single-cell transcriptomics of mouse foregut organogenesis. Building on established methods, hPSC-derived lateral plate mesoderm (LPM) treated with either retinoic acid (RA) or RA together with a Hedgehog (HH) agonist generates posterior or anterior foregut splanchnic mesoderm respectively (pFG-SpM or aFG-SpM) after 4 days cultures. These are directed into organ-specific mesenchyme lineages by the combinatorial activation or inhibition of WNT, BMP, RA or HH pathways from days 4 to 7 in cultures. By day 7 the cultures are enriched for different types of mesoderm with distinct molecular signatures: 60–90% pure liver septum transversum/mesothelium-like (STM/mesothelium), 70–80% pure liver-like fibroblasts (LF), and populations of ~35% respiratory-like mesoderm (RM), gastric-like mesoderm (GM), or esophageal-like mesoderm (EM). This protocol can be performed by anyone with moderate experience differentiating hPSCs and provides a novel platform to study human mesoderm development and can be used to engineer more complex foregut tissue for disease modeling and regenerative medicine.
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影响因子：
21.3
作者：
Chen YW;Huang SX;de Carvalho ALRT;Ho SH;Islam MN;Volpi S;Notarangelo LD;Ciancanelli M;Casanova JL;Bhattacharya J;Liang AF;Palermo LM;Porotto M;Moscona A;Snoeck HW
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Snoeck HW
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通讯作者：
Muensterberg, Andrea
影响因子：
2.7
作者：
Edwards NA;Shacham-Silverberg V;Weitz L;Kingma PS;Shen Y;Wells JM;Chung WK;Zorn AM
通讯作者：
Zorn AM
影响因子：
9.8
作者：
Tang, Xing;Liu, Huayang;Leone, Gustavo
通讯作者：
Leone, Gustavo
影响因子：
7.7
作者：
Dye BR;Hill DR;Ferguson MA;Tsai YH;Nagy MS;Dyal R;Wells JM;Mayhew CN;Nattiv R;Klein OD;White ES;Deutsch GH;Spence JR
通讯作者：
Spence JR