Alteration of the Chemotactic Response of Human Skin Fibroblasts to PDGF by Growth Factors
Alteration of the Chemotactic Response of Human Skin Fibroblasts to PDGF by Growth Factors
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生长因子改变人皮肤成纤维细胞对 PDGF 的趋化反应
DOI：
10.1111/j.1346-8138.1992.tb03752.x
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Yasumasa Ishibashi
中科院分区：
文献类型：
--
作者：
Y. Soma;K. Takehara;Yasumasa Ishibashi
健康成人皮肤的植物，并在补充有10%胎牛血清（FBS）的最低必需培养基（MEM）中培养。细胞在第三次传代前用于实验。生长因子PDGF和TGF-β购自R&D Inc. EGF和碱性FGF购自Sigma Chemical Company。趋化性测定通过使用如前所述的盲孔室进行趋化性测定（8）。将供试品在含0.2 mg/ml BSA的MEM中稀释，并加入每个盲孔室的下孔中。然后用胶原蛋白包被的聚碳酸酯过滤器（Nucleopore，8微米直径的孔）覆盖该较低的孔。然后将上孔固定在适当位置，并将从组织培养瓶中新鲜释放的细胞加入到含有2 mg/ml BSA的MEM中。在37°C下在90%空气和10%C02中孵育4小时后，移除过滤器，固定细胞并通过Diff-Quick染色进行染色。材料和方法细胞培养人皮肤成纤维细胞从体外培养
plants of healthy human adult skin and cultured in minimal essential medium (MEM) supplemented with 10% fetal bovine serum (FBS). Cells were used for the experiments prior to the third passage. Growth Factors PDGF and TGF-beta were purchased from R&D Inc. EGF and basic FGF were purchased from Sigma Chemical Company. Chemotaxis Assay The chemotaxis assay was performed by using blind well chambers as described previously (8). The test substance was diluted in MEM containing 0.2 mg/ml BSA and added to the lower well of each blind well chamber. This lower well was than covered with a collagen-coated polycarbonate filter (Nucleopore, 8 micrometer-diameter pores). The upper well was then fixed in place, and cells freshly released from tissue culture flasks were added to it in MEM containing 2 mg/ml BSA. After 4 hours of incubation at 37°C in 90% air and 10% CO2, the filters were removed and the cells were fixed and stained by Diff-Quick stain. The upper surface cells Materials and Methods Cell Culture Human skin fibroblasts were grown from exIntroduction