Differentiation of Human Intestinal Organoids with Endogenous Vascular Endothelial Cells
Differentiation of Human Intestinal Organoids with Endogenous Vascular Endothelial Cells
复制标题
DOI：
10.1016/j.devcel.2020.07.023
复制
发表时间：
2020-08-24
影响因子：
11.8
通讯作者：
Spence, Jason R.
中科院分区：
文献类型：
--
作者：
Holloway, Emily M.;Wu, Joshua H.;Spence, Jason R.
人多能干细胞（hPSC）衍生的肠类器官（HIO）缺乏在天然器官（包括脉管系统）中发现的一些细胞群体。使用单细胞RNA测序（scRNA-seq），我们已经鉴定了在HIO分化早期存在的内皮细胞（EC）群体，其在培养物中随着时间的推移而下降。在这里，我们开发了一种方法来扩大和维持这种内源性的细胞内的HIO（vHIO）。鉴于EC具有器官特异性基因表达、形态和功能，我们使用批量RNA-seq和scRNA-seq来询问发育中的人肠、肺和肾，以鉴定器官富集的EC基因特征。通过将这些基因签名和经验证的标志物与HIO EC进行比较，我们发现体外生长的HIO EC与天然肠EC相对于肾和肺具有最高的相似性。总之，这些数据证明HIO可以共分化天然EC群体，所述天然EC群体在体外用精氨酸特异性EC转录签名适当地模式化。
Human pluripotent stem cell (hPSC)-derived intestinal organoids (HIOs) lack some cellular populations found in the native organ, including vasculature. Using single-cell RNA sequencing (scRNA-seq), we have identified a population of endothelial cells (ECs) present early in HIO differentiation that declines over time in culture. Here, we developed a method to expand and maintain this endogenous population of ECs within HIOs (vHIOs). Given that ECs possess organ-specific gene expression, morphology, and function, we used bulk RNA-seq and scRNA-seq to interrogate the developing human intestine, lung, and kidney in order to identify organ-enriched EC gene signatures. By comparing these gene signatures and validated markers to HIO ECs, we find that HIO ECs grown in vitro share the highest similarity with native intestinal ECs relative to kidney and lung, Together, these data demonstrate that HIOs can co-differentiate a native EC population that is properly patterned with an intestine-specific EC transcriptional signature in vitro.