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孕中期羊水来源的培养细胞:IV. 通过克隆全细胞匀浆的一维和二维电泳进行细胞类型鉴定

Cultivated cells from mid-trimester amniotic fluids: IV. Cell type indentification via one and two-dimensional electrophoresis of clonal whole cell homogenates.

作者信息

Johnston P, Salk D, Martin G M, Hoehn H

出版信息

Prenat Diagn. 1982 Apr;2(2):79-88. doi: 10.1002/pd.1970020202.

Abstract

Clones of cultivated amniotic fluid cells that have distinct morphologic and growth characteristics (F, AF and E-type) were examined by one-dimensional SDS polyacrylamide gel electrophoresis (SDS-PAGE) and by two-dimensional electrophoresis employing isoelectric focusing and SDS-PAGE (IEF-PAGE). No qualitative differences in band pattern were observed in SDS-PAGE between the various amniotic fluid cell types, but consistent quantitative differences in the ratios of four bands of presumed filamentous proteins provided good distinction between amniotic fluid cells and postnatal skin fibroblast-like cells. By adding separation on the basis of electrical charge to that of molecular size (IEF-PAGE), we observed reproducible qualitative differences in the protein spot patterns between F and both AF and E-type amniotic fluid cells. At least eight discrete proteins appear not be synthesized by prenatal F-type cells in comparison with their isogenic AF and E counterparts under identical culture conditions. The two-dimensional electrophoretic patterns thus confirm that F and AF amniotic cells, in spite of their morphologic and growth kinetic similarities, are developmentally distinct cell types that retain their differentiated states in culture.

摘要

通过一维十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)以及采用等电聚焦和SDS-PAGE的二维电泳(IEF-PAGE),对具有不同形态学和生长特征(F型、AF型和E型)的培养羊水细胞克隆进行了检测。在SDS-PAGE中,不同类型羊水细胞的条带模式未观察到定性差异,但推测为丝状蛋白的四条带的比例存在一致的定量差异,这为羊水细胞和产后皮肤成纤维细胞样细胞提供了良好的区分。通过在分子大小分离的基础上增加基于电荷的分离(IEF-PAGE),我们观察到F型与AF型和E型羊水细胞之间在蛋白质斑点模式上存在可重复的定性差异。与在相同培养条件下的同基因AF型和E型对应细胞相比,至少有八种离散蛋白似乎不是由产前F型细胞合成的。二维电泳模式因此证实,F型和AF型羊水细胞尽管在形态学和生长动力学上相似,但它们是发育上不同的细胞类型,在培养中保持其分化状态。

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