INITIAL CULTURE BEHAVIOR OF RAT BLASTOCYSTS ON SELECTED FEEDER CELL-LINES
INITIAL CULTURE BEHAVIOR OF RAT BLASTOCYSTS ON SELECTED FEEDER CELL-LINES
复制标题
DOI：
10.1002/mrd.1080400307
复制
发表时间：
1995-03-01
影响因子：
2.5
通讯作者：
CLARKE, NJ
中科院分区：
文献类型：
--
作者：
OUHIBI, N;SULLIVAN, NF;CLARKE, NJ
为了加深对大鼠胚胎培养的了解，并尝试分离囊胚来源的细胞系，我们研究了四个品系的大鼠在四个不同饲养层上的囊胚的初始生长行为。饲养层为小鼠胚胎成纤维细胞(STO)、原代小鼠(MEF)或大鼠(REF)胚胎成纤维细胞连续细胞系和大鼠子宫上皮细胞(RUCs)连续细胞系。在大鼠胚胎培养中使用了一种对小鼠胚胎干细胞具有最佳培养效率的培养液。每种培养系统都允许囊胚孵化和附着，也就是说，在培养的头2天，每个饲养器和每个品系的行为都是相似的。随后，成纤维细胞饲养层(STO>；MEF>；REF)上的内细胞团(ICM)细胞迅速分化，并在原代培养3-6天后基本完成。在RUC上，发现ICM的大小增加而没有分化，直到并包括第4天，在某些情况下更长。在REF和RUC饲养层上解聚和传代ICM，获得胚胎来源细胞。REF上可见圆形、屈光性和上皮样细胞，RUC上可见ES样细胞集落。ES样细胞表达碱性磷酸酶和阶段特异性胚胎抗原1。这是向分化和培养大鼠多能ES细胞迈出的重要的第一步。(C)1995年Wiley-Liss公司
To increase our understanding of rat embryos in culture and to attempt the isolation of blastocyst-derived cell lines, we examinated the initial growth behaviour of rat blastocysts from four strains of rat on four different feeder cell layers. The feeders used were a continuous cell line of murine embryonic fibroblasts (STO), primary mouse (MEF) or primary rat (REF) embryonic fibroblasts, and a continuous cell line of rat uterine epithelial cells (RUCs). A medium that gave optimum plating efficiencies for murine ES cells was used in the rat embryo culture. Each culture system allowed hatching and attachment of the blastocysts, that is, the behaviour was similar on each feeder and each strain for the first 2 days in culture. Subsequently, there was a rapid differentiation of the Inner Cell Mass (ICM) cells on fibroblastic feeder cell layers (STO > MEF > REF), and this was generally complete after 3-6 days in primary culture. On RUCs, the ICM was found to increase in size without differentiation up to and including day 4 and in some cases longer. Embryo-derived cells were obtained by disaggregating and passaging ICMs on REF and RUC feeders. Rounded, refractile, and epithelial-like cells were isolated on REF and colonies of ES-like cells on the RUCs. The ES-like cells were positive for expression of alkaline phosphatase and stage-specific embryonic-antigen 1. This is an important first step towards the derivation and culture of pluripotent ES cells from the rat. (C) 1995 Wiley-Liss, Inc.