シグナル経路制御によるヒトES/iPS細胞からの肺胞細胞の誘導

通过控制信号通路从人 ES/iPS 细胞诱导肺泡细胞

• 批准号: 26670417
• 负责人: シャーサバラニ ホセイン
• 金额: $ 2.33万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2014
• 资助国家: 日本
• 起止时间: 2014-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-26670417/
• 关键词: lung differentiation; hPSCs; Signal transduction

In an attempt to direct differentiation of hPSCs to the lung alveolar cells which are not restricted by technical considerations for the clinical application, various inhibitors and microenvironment factors which expected to participate in this developmental process have been assessed in current study based on developmental paradigms,We first tried to culture hPSCs with lung differentiation basal medium to make embryoid bodies. To enhance differentiation, three kinds of inhibitors (Wnt, BMP and ROCK). After further culturing the cells with inhibitors with growth factors (bFGF, Activin A) and TGFb inhibitors. Specification of the single cells of embryoid bodies to lung progenitor cells investigated using long culturing of single cells different concentration of various signal inhibitors including GSK3 inhibitors (Kenpaullone and CHIR99021), RA and cocktail of growth factors positively effect lung differentiation (FGF10, FGF7, BMP4, EGF) on fibronectin-coated plates, to probe the instructive mechanisms of endodermal lung differentiation.Because of the lack of time and terminating the project in its mid-stage, unfortunately we could not succeed to efficiently generate alveolar cells from hPSCs through evaluation of all possible signaling mechanisms that expected to regulate lung specification. Thus further study for elucidate other critical pathways in lung differentiation must used to understand the mechanisms of lung cells differentiation.However, a review paper has been prepared entitled ”Insights into the directed differentiation of hPSCs to lung differentiation”.

为了使hPSC分化为不受临床应用技术限制的肺泡细胞，本研究基于发育范式评估了预期参与该发育过程的各种抑制剂和微环境因素。我们首先尝试用肺分化基础培养基培养hPSC以制备胚状体。为了促进分化，三种抑制剂（Wnt，BMP和ROCK）。在用生长因子抑制剂（bFGF、激活素A）和TGF β抑制剂进一步培养细胞后。使用不同浓度的各种信号抑制剂（包括GSK 3抑制剂）的单细胞长期培养研究胚状体的单细胞向肺祖细胞的特化（Kenpaullone和CHIR99021）、RA和生长因子混合物积极影响肺分化（FGF 10、FGF 7、BMP 4、EGF）在纤连蛋白包被的板上，探讨内胚层肺分化的指导机制，由于时间紧迫，项目中途终止，不幸的是，我们不能通过评估预期调节肺特化的所有可能的信号传导机制成功地从hPSC有效地产生肺泡细胞。因此，进一步研究阐明肺分化中的其他关键途径必须用于了解肺细胞分化的机制。然而，已经准备了题为“Insights into the directed differentiation of hPSCs to lung differentiation”的综述文章。