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使用三明治技术在I型胶原蛋白中培养的成纤维细胞的行为。

Behaviour of fibroblastic cells cultured in collagen I using the sandwich technique.

作者信息

Haas V R, Santos A R, Wada M L

机构信息

Department of Cell Biology, Institute of Biology, State University of Campinas, UNICAMP, SP, Brasil.

出版信息

Cytobios. 2001;106 Suppl 2:255-67.

Abstract

Vero cells, a fibroblastic lineage derived from fibroblastic kidney cells of the African green monkey, were cultivated by means of the sandwich technique, involving glass coverslip/collagen and collagen/collagen with varied foetal calf serum concentrations in the culture medium. The cells, cultured on coverslips, then received a type I collagen gel layer on top, and migrated from the coverslip to the collagen layer. When the cells were cultivated on collagen followed by a covering of type I collagen gel, the cells migrated into both collagen layers. Cellular morphology was similar, independent of the type of sandwich and serum concentration used. Cells in contact with collagen either migrated into the layer or formed a basal lamina separating them from the collagen matrix. The formation of a basal lamina with laminin and collagen IV deposition was most noticeable when 20% foetal calf serum was used in the culture medium. Cellular infiltration into the collagen gel was most evident with the use of 10% foetal calf serum. The gel contraction was similar for the two serum concentrations employed.

摘要

Vero细胞源自非洲绿猴的成纤维细胞性肾细胞,采用三明治技术培养,该技术涉及在培养基中使用不同胎牛血清浓度的玻璃盖玻片/胶原蛋白和胶原蛋白/胶原蛋白组合。将细胞培养在盖玻片上,然后在顶部覆盖一层I型胶原蛋白凝胶,细胞从盖玻片迁移到胶原蛋白层。当细胞在胶原蛋白上培养,随后覆盖I型胶原蛋白凝胶时,细胞迁移到两层胶原蛋白中。细胞形态相似,与所使用的三明治类型和血清浓度无关。与胶原蛋白接触的细胞要么迁移到该层中,要么形成将它们与胶原蛋白基质分隔开的基膜。当在培养基中使用20%胎牛血清时,带有层粘连蛋白和IV型胶原蛋白沉积的基膜形成最为明显。使用10%胎牛血清时,细胞向胶原蛋白凝胶中的浸润最为明显。对于所采用的两种血清浓度,凝胶收缩情况相似。

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