Differentiation of human embryonic stem cells into embryoid bodies comprising the three embryonic germ layers
Differentiation of human embryonic stem cells into embryoid bodies comprising the three embryonic germ layers
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DOI：
10.1007/bf03401776
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发表时间：
2000-02-01
影响因子：
5.7
通讯作者：
Benvenisty, N
中科院分区：
文献类型：
--
作者：
Itskovitz-Eldor, J;Schuldiner, M;Benvenisty, N
背景：胚胎干细胞（ES细胞）是从囊胚中分离的细胞系。小鼠ES细胞被证明是真正的多能细胞，因为它们分化成所有的胚胎谱系。然而，恒河猴ES细胞的体外分化有些不一致和混乱。材料与方法：人ES细胞悬浮培养，诱导其分化为胚状体（EBs）。利用逆转录聚合酶链反应（RT-PCR）和原位杂交技术，观察了ES细胞和EB细胞的分化状态，并对ES细胞的分化进行了研究。我们的研究结果表明，诱导表达的细胞特异性基因的人ES细胞分化成EB。在人类EB中，我们可以显示出不同细胞谱系的胚胎标志物的特征性区域表达，即zeta-珠蛋白（中胚层），神经丝68Kd（外胚层）和甲胎蛋白（内胚层）。此外，我们提出了一个同步脉冲胚状体，表达心肌标记物α-心脏肌动蛋白。此外，解离的胚状体和平板细胞单层的结果在多种形态，其中细胞与神经元的外观，表达神经丝68KD chains.Conclusion：人ES细胞可以reproducibly分化成EB包括三个胚胎胚层体外。诱导形成含有神经元、造血和心脏来源的细胞的人胚状体的能力将在研究早期人胚胎发育以及移植医学中有用。
Background: Embryonic stem (ES) cells are lines of cells that are isolated from blastocysts. The murine ES cells were demonstrated to be true pluripotent cells as they differentiate into all embryonic lineages. Yet, in vitro differentiation of rhesus ES cells was somewhat inconsistent and disorganized. The recent isolation of human ES cells calls for exploring their pluripotential nature.Materials and Methods: Human ES cells were grown in suspension to induce their differentiation into embryoid bodies (EBs). The differentiation status of the human ES cells and EBs was analyzed by following the expression pattern of several lineage-specific molecular markers using reverse transcription polymerase chain reaction (RT-PCR) and in situ hybridization.Results: Here we report the induction in vitro of cystic embryoid bodies from human ES cells. Our findings demonstrate induction of expression of cell-specific genes during differentiation of the human ES cells into EBs. In the human EBs, we could show a characteristic regional expression of embryonic markers specific to different cellular lineages, namely, zeta-globin (mesoderm), neurofilament 68Kd (ectoderm), and alpha-fetoprotein (endoderm). Moreover, we present a synchronously pulsing embryoid body that expresses the myocardium marker a-cardiac actin. In addition, dissociating the embryoid bodies and plating the cells as monolayers results in multiple morphologies, among them cells with neuronal appearance that express neurofilament 68Kd chain.Conclusion: Human ES cells can reproducibly differentiate in vitro into EBs comprising the three embryonic germ layers. The ability to induce formation of human embryoid bodies that contain cells of neuronal, hematopoietic and cardiac origins will be useful in studying early human embryonic development as well as in transplantation medicine.