微流体羊膜囊胚状体(μPASE)是人类多能干细胞(hPSC)衍生的多细胞人类胚胎样结构,具有类似于植入后早期人类胚胎囊的逐渐发育的分子和形态特征。微流体装置专门设计用于控制hPSC簇的形成,并将簇暴露于不同的形态发生原环境,允许μPASEs在高度可控的情况下发展,可重复,和可扩展的时尚。μPASE模型显示了人类胚胎发育标志,例如上胚层的腔形成,羊膜腔形成,以及原始生殖细胞和胃泌素细胞(或中内胚层细胞)的规格。这里,我们提供复制μPASE所需的详细说明,包括用于表征在不同实验条件下获得的μPASEs的免疫荧光染色和细胞检索方案。
The microfluidic amniotic sac embryoid (μPASE) is a human pluripotent stem cell (hPSC)-derived multicellular human embryo-like structure with molecular and morphological features resembling the progressive development of the early post-implantation human embryonic sac. The microfluidic device is specifically designed to control the formation of hPSC clusters and expose the clusters to different morphogen environments, allowing the development of μPASEs in a highly controllable, reproducible, and scalable fashion. The μPASE model displays human embryonic developmental landmarks such as lumenogenesis of the epiblast, amniotic cavity formation, and the specification of primordial germ cells and gastrulating cells (or mesendoderm cells). Here, we provide detailed instructions needed to reproduce μPASEs, including the immunofluorescence staining and cell retrieval protocols for characterizing μPASEs obtained under different experimental conditions.