BMP4-directed trophoblast differentiation of human embryonic stem cells is mediated through ΔNp63+ cytotrophoblast stem cell state
BMP4-directed trophoblast differentiation of human embryonic stem cells is mediated through ΔNp63+ cytotrophoblast stem cell state
复制标题
DOI：
10.1242/dev.092155
复制
发表时间：
2013-10-01
期刊：
影响因子：
4.6
通讯作者：
Parast, Mana M.
中科院分区：
文献类型：
--
作者：
Li, Yingchun;Moretto-Zita, Matteo;Parast, Mana M.
胎盘是适当胎儿发育所必需的瞬态器官。它的主要功能成分是滋养细胞，它源自外胚外外胚层。由于缺乏适当的体外模型系统，人们对人类胚胎的早期滋养细胞分化知之甚少。在存在喂食器条件培养基的情况下，BMP4处理后，人类胚胎干细胞（HESC）分化为功能性滋养细胞。但是，由于缺乏滋养细胞祖细胞种群的证据，该模型并未被广泛接受。我们先前已经表明，p63是p53核蛋白家族的成员，在增生性细胞增生性细胞（CTB）中表达，是为植入部位的绒毛膜绒毛的终末分化的合成细胞细胞（STB）的前体。在这里，我们表明，通过与原代人胎盘组织进行直接比较，通过基因表达分析直接比较BMP4处理的hESC将分化为真正的CTB。我们表明，在初级CTB中，随着细胞分化为STB，p63水平降低，并且强迫p63的表达保持细胞周期蛋白B1并抑制STB分化。我们还确定，与体内事件类似，hESC分化为滋养层的特征是p63（+）/krt7（+）CTB干细胞状态，然后形成功能性KLF4（+）STB和HLA-G（+） evt。最后，我们说明shRNA对p63的下调会抑制hESC分化为功能性滋养细胞。综上所述，我们的结果表明，经BMP4处理的hESC是人类滋养细胞分化的极好模型，密切模仿了体内从p63（+）CTB干细胞到终止分化的滋养细胞亚型的体内进展。
The placenta is a transient organ that is necessary for proper fetal development. Its main functional component is the trophoblast, which is derived from extra-embryonic ectoderm. Little is known about early trophoblast differentiation in the human embryo, owing to lack of a proper in vitro model system. Human embryonic stem cells (hESCs) differentiate into functional trophoblast following BMP4 treatment in the presence of feeder-conditioned media; however, this model has not been widely accepted, in part owing to a lack of proof for a trophoblast progenitor population. We have previously shown that p63, a member of the p53 family of nuclear proteins, is expressed in proliferative cytotrophoblast (CTB), precursors to terminally differentiated syncytiotrophoblast (STB) in chorionic villi and extravillous trophoblast (EVT) at the implantation site. Here, we show that BMP4-treated hESCs differentiate into bona fide CTB by direct comparison with primary human placental tissues and isolated CTB through gene expression profiling. We show that, in primary CTB, p63 levels are reduced as cells differentiate into STB, and that forced expression of p63 maintains cyclin B1 and inhibits STB differentiation. We also establish that, similar to in vivo events, hESC differentiation into trophoblast is characterized by a p63(+)/KRT7(+) CTB stem cell state, followed by formation of functional KLF4(+) STB and HLA-G(+) EVT. Finally, we illustrate that downregulation of p63 by shRNA inhibits differentiation of hESCs into functional trophoblast. Taken together, our results establish that BMP4-treated hESCs are an excellent model of human trophoblast differentiation, closely mimicking the in vivo progression from p63(+) CTB stem cells to terminally differentiated trophoblast subtypes.