Lung epithelial cell specification in human pluripotent stem cells

肺移植是治疗终末期肺部疾病的唯一权威疗法。 因严重并发症和捐赠器官短缺而受阻。一种潜力 至少在某些上皮细胞受到特殊影响的疾病中，替代疗法是STEM 细胞或细胞替代疗法。这种疗法的应用还很遥远， 然而。细胞治疗的先决条件是无限量供应正确的上皮细胞。 祖细胞和有效的植入程序。在这份提案中，我们主要关注 第一个目标，并使用通用损伤模型来证明这些细胞的潜力。 为了解决潜在的呼吸道前体细胞无限供应的问题，我们将重点放在了 胚胎干细胞(ESCs)和诱导多能干细胞(IPSCs) 被称为人类多能干细胞(HPSCs)。我们在这里描述了从 一群可能是胎肺末端远端祖细胞(PDTP)的hPSCs 持续扩展(截至目前18个月)，并高效地重新填充大部分或全部 免疫缺陷NSG小鼠博莱霉素损伤肺中的谱系。此外，这些 细胞可以转化为具有基底细胞(BCS)所有特征的细胞，干细胞 然而，呼吸道细胞表现出更有限的扩张。尽管事实是 PDTPs植入博莱霉素处理的NSG小鼠的呼吸道，其来源的BC样细胞 没有植入，这表明BCS可能没有植入能力或 博莱霉素不是这些细胞的合适损伤模型。这项提议的目标是 对这两个细胞群体进行分子表征，并评估它们在各种 损伤模型。这项建议的具体目标是： 目的1：pDTPs和BC样细胞的分子和体外特性 目的2.hPSC来源的pDTPs和BC样细胞的体内潜能。 目的3.pDTPs的培养条件及克隆性。

Lung transplantation, the only definitive treatment for end-stage lung disease, remains hampered by severe complications and a shortage of donor organs. One potential alternative for at least some diseases where the epithelium is specifically affected is stem cell or cellular replacement therapy. Application of this type of therapy is still far away, however. Prerequisites for cellular therapy are an unlimited supply of the correct epithelial progenitors and an efficient engraftment procedure. In this proposal, we focus primarily on the first goal, and use generic injury models to demonstrate the potential of these cells. To address the issue of an unlimited supply of potential airway progenitors, we focused on embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), collectively termed human pluripotent stem cells (hPSCs). We describe here the generation from hPSCs of a population of putative fetal lung distal tip progenitors (pDTPs) that can be expanded continuously (up to now 18 months) and efficiently repopulates most or all lineages in bleomycin-injured lungs of immunodeficient NSG mice. Furthermore, these cells can be converted to cells with all the characteristics of basal cells (BCs), the stem cells of the airway, which, however, showed more limited expansion. Despite the fact the pDTPs engrafted in the airways of bleomycin-treated NSG mice, the derived BC-like cells did not engraft, suggesting that BCs may not possess engraftment capacity or that bleomycin is not an appropriate injury model for these cells. The goal of this proposal is to molecularly characterize both cell populations and evaluate their potentials in a variety of injury models. The specific aims of the proposal are: Aim 1: Molecular and in vitro characterization of pDTPs and BC-like cells Aim 2. In vivo potential of hPSC-derived pDTPs and BC-like cells. Aim 3. Culture conditions and clonality of pDTPs.