Simultaneous differentiation of endothelial and trophoblastic cells derived from human embryonic stem cells
Simultaneous differentiation of endothelial and trophoblastic cells derived from human embryonic stem cells
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DOI：
10.1089/scd.2006.0013
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发表时间：
2007-06-01
影响因子：
4
通讯作者：
Hatzfeld, Jacques A.
中科院分区：
文献类型：
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作者：
Peiffer, Isabelle;Belhomme, Denis;Hatzfeld, Jacques A.
在这里，我们提出了一个简单的两步体外模型，来自人胚胎干（hES）细胞的血管化滋养层组织。第一步是在半固体甲基纤维素培养基中悬浮形成囊性胚状体 (EB)，其中内皮血小板/内皮细胞粘附分子-1 (PECAM-1(+)) 细胞网络发育。第二步，这些 EB 沉积在未经处理的聚苯乙烯组织培养板的底部，通过 EB 的离心生长导致贴壁细胞层的出现，与 PECAM-1+ 网络相关。该粘附层的细胞表达 VE-钙粘蛋白 (CD144)、PECAM-1 (CD31) 和甲胎蛋白 (alpha-FP)。 ss-人绒毛膜促性腺激素 (ss-hCG) 的分泌以及细胞滋养层和合体滋养层标记 GB25 的存在强烈提示滋养层分化。与作为参考的 hES 细胞基因表达相比，INSL4 基因（细胞和合体滋养层标记物）以及其他滋养层基因（例如 CGA、CDX1、CDX2 和 HAND1）也在贴壁层中高度表达。相反，自我更新基因，如 TERT、POU5F1、ZFP42、GDF3 和 NODAL 的表达下降。没有外胚层或内胚层基因表达，但中胚层基因 PECAM-1 和 GATA2 表达。在第二步中去除 EB 的可能性将允许分析与滋养层粘附层相比，它们对血管生成或可能的成血管细胞形成的相对贡献。这种原始的血管滋养层模型还可以提供研究正常和病理性胎盘发育早期步骤的工具。
Here we present a simple two-step in vitro model of vascularized trophoblastic tissue derived from human embryonic stem (hES) cells. The first step is the formation of cystic embryoid bodies (EBs) in suspension in a semisolid methyl cellulose medium, within which an endothelial platelet/endothelial cell adhesion molecule-1 (PECAM-1(+)) cell network develops. In a second step, deposition of these EBs on the bottom of nontreated, polystyrene tissue culture plates, leads by centrifugal outgrowth of the EB to the emergence of an adherent cell layer, with which a PECAM-1+ network is associated. Cells of this adherent layer expressed VE-cadherin (CD144), PECAM-1 (CD31), and alpha-fetoprotein (alpha-FP). Trophoblastic differentiation was strongly suggested by the secretion of ss-human chorionic gonadotropin (ss-hCG) and by the presence of the cytotrophoblast and syncytiotrophoblast marker GB25. The INSL4 gene, a cyto- and syncytio-trophoblast marker, was also highly expressed in the adherent layer, as well as other trophoblast genes such as CGA, CDX1, CDX2, and HAND1, compared to hES cell gene expression taken as reference. In contrast, expression of self-renewal genes, such as TERT, POU5F1, ZFP42, GDF3, and NODAL were decreased. No ectodermal or endodermal genes were expressed, but the mesodermal genes PECAM-1 and GATA2 were. The possibility of removing the EBs during the second step would permit analysis of their relative contribution to angiogenesis or possible hemangioblast formation, compared to that of the trophoblastic adherent layer. This primitive vascularized trophoblastic model could also provide a tool to study early steps of normal and pathological placental development.