Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800, Australia.
Murdoch Childrens Research Institute, The Royal Children's Hospital, Parkville, VIC 3052, Australia.
Stem Cell Reports. 2014 May 22;2(6):925-37. doi: 10.1016/j.stemcr.2014.04.009. eCollection 2014 Jun 3.
Thymic epithelial cells (TECs) play a critical role in T cell maturation and tolerance induction. The generation of TECs from in vitro differentiation of human pluripotent stem cells (PSCs) provides a platform on which to study the mechanisms of this interaction and has implications for immune reconstitution. To facilitate analysis of PSC-derived TECs, we generated hESC reporter lines in which sequences encoding GFP were targeted to FOXN1, a gene required for TEC development. Using this FOXN1 (GFP/w) line as a readout, we developed a reproducible protocol for generating FOXN1-GFP(+) thymic endoderm cells. Transcriptional profiling and flow cytometry identified integrin-β4 (ITGB4, CD104) and HLA-DR as markers that could be used in combination with EpCAM to selectively purify FOXN1(+) TEC progenitors from differentiating cultures of unmanipulated PSCs. Human FOXN1(+) TEC progenitors generated from PSCs facilitate the study of thymus biology and are a valuable resource for future applications in regenerative medicine.
胸腺上皮细胞 (TECs) 在 T 细胞成熟和诱导耐受中发挥着关键作用。从人多能干细胞 (PSCs) 的体外分化中产生 TECs，为研究这种相互作用的机制提供了一个平台，并对免疫重建具有重要意义。为了便于分析 PSC 衍生的 TECs，我们生成了 hESC 报告细胞系，其中编码 GFP 的序列靶向 FOXN1，这是 TEC 发育所必需的基因。我们使用这个 FOXN1 (GFP/w) 细胞系作为读出信号，开发了一种可重复的方案，用于生成 FOXN1-GFP(+) 胸腺内胚层细胞。转录组分析和流式细胞术鉴定了整合素-β4 (ITGB4，CD104) 和 HLA-DR，这些标志物可以与 EpCAM 结合，从未经过处理的 PSCs 分化培养物中选择性地纯化 FOXN1(+) TEC 祖细胞。从 PSCs 产生的人 FOXN1(+) TEC 祖细胞促进了对胸腺生物学的研究，并且是再生医学中未来应用的有价值资源。
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