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[培养细胞胞外基质的分离与特性]

[Isolation and characteristics of the extracellular matrix of cultured cells].

作者信息

Domnina L V, Liubimov A V

出版信息

Tsitologiia. 1988 Mar;30(3):299-304.

PMID:3046077
Abstract

Sodium deoxycholate extraction was used to isolate extracellular matrix from various cultured cells: human and murine embryonic fibroblasts, epithelial lines of mouse (MPTR), rat (IAR 2 and IAR 20), pig (SPEV) and cow (FBT). Protein composition of the matrix was studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunofluorescence. The matrix morphology was investigated by scanning electron microscopy. In cell lines FBT and MPTR the major component of the matrix was laminin, whereas in other lines and fibroblasts it was fibronectin. The matrix of the majority of lines had a fibrillar structure, and the fibrils usually formed networks. MPTR cells had a punctate matrix composed of laminin and collagen type IV, densely covering the substratum. The treatment of the matrix by hyaluronidase and/or DNAase I did not influence its protein composition. The isolated matrix of different structure and composition may serve a biological substratum in studies of normal tumor cell behavior in tissue culture.

摘要

使用脱氧胆酸钠提取法从多种培养细胞中分离细胞外基质

人和鼠胚胎成纤维细胞、小鼠上皮细胞系(MPTR)、大鼠上皮细胞系(IAR 2和IAR 20)、猪上皮细胞系(SPEV)和牛上皮细胞系(FBT)。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳和免疫荧光研究基质的蛋白质组成。通过扫描电子显微镜研究基质形态。在FBT和MPTR细胞系中,基质的主要成分是层粘连蛋白,而在其他细胞系和成纤维细胞中是纤连蛋白。大多数细胞系的基质具有纤维状结构,且纤维通常形成网络。MPTR细胞具有由层粘连蛋白和IV型胶原组成的点状基质,密集覆盖在基质上。用透明质酸酶和/或DNA酶I处理基质不影响其蛋白质组成。分离得到的具有不同结构和组成的基质可作为组织培养中正常肿瘤细胞行为研究的生物基质。

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