Generation of 3D Trophoblast Organoids from Human Naïve Pluripotent Stem Cells.
Generation of 3D Trophoblast Organoids from Human Naïve Pluripotent Stem Cells.
复制标题
从人类幼稚多能干细胞生成 3D 滋养层类器官。
DOI：
10.1007/7651_2023_496
复制
发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Theunissen,ThoroldW
中科院分区：
文献类型：
--
作者：
Karvas,RowanM;Theunissen,ThoroldW
人类胎盘是一个短暂的器官，其功能是在整个妊娠期间支持胎儿的需要。滋养层细胞是胎盘内发现的主要上皮细胞，并且包括在胎儿-母体通信中具有专门作用的各种不同的细胞类型。我们对人类滋养层发育的理解仍然有限，这是由于获取妊娠早期胎盘组织的伦理和法律的限制，以及普通动物模型无法复制灵长类胎盘发育。因此，重要的是要推进在体外模型的人滋养层发育的基础上，研究妊娠相关的并发症和疾病。在本章中，我们描述了一种从幼稚的人类多能干细胞（hPSC）生成3D滋养层类器官的方案。由此产生的干细胞衍生的滋养层类器官（SC-TO）包含不同的细胞滋养层（CTB）、合胞体滋养层（STB）和绒毛外滋养层（EVT）细胞类型，这些细胞类型与人类植入后胚胎中的滋养层身份密切对应。我们讨论了通过免疫荧光、流式细胞术、mRNA和microRNA表达谱以及胎盘激素分泌来表征SC-TO的方法。此外，SC-TO可以分化成专门的3D EVT类器官，当与人子宫内膜细胞共培养时，其显示出强大的侵袭。因此，本文所述的方案提供了人类胎盘发育和滋养层侵袭的可访问的3D模型系统。
The human placenta is a transient organ that functions to support the needs of the fetus throughout gestation. Trophoblasts are the major epithelial cells found within the placenta and comprise a variety of distinct cell types with specialized roles in fetal-maternal communication. Our understanding of human trophoblast development remains limited due to ethical and legal restrictions on accessing first-trimester placental tissues, as well as the inability of common animal models to replicate primate placental development. It is therefore important to advance in vitro models of human trophoblast development as a basis for studying pregnancy-associated complications and diseases. In this chapter, we describe a protocol for generating 3D trophoblast organoids from naïve human pluripotent stem cells (hPSCs). The resulting stem-cell-derived trophoblast organoids (SC-TOs) contain distinct cytotrophoblast (CTB), syncytiotrophoblast (STB), and extravillous trophoblast (EVT) cell types, which closely correspond to trophoblast identities in the human post-implantation embryo. We discuss methods for characterizing SC-TOs by immunofluorescence, flow cytometry, mRNA and microRNA expression profiling, and placental hormone secretion. Furthermore, SC-TOs can undergo differentiation into specialized 3D EVT organoids, which display robust invasion when co-cultured with human endometrial cells. Thus, the protocol described herein offers an accessible 3D model system of human placental development and trophoblast invasion.