The effect of the dimeric and multimeric forms of fibronectin on the adhesion and growth of primary glomerular cells.
The effect of the dimeric and multimeric forms of fibronectin on the adhesion and growth of primary glomerular cells.
复制标题
纤连蛋白二聚体和多聚体形式对原代肾小球细胞粘附和生长的影响。
DOI：
10.1016/0014-4827(83)90005-8
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发表时间：
1983
影响因子：
3.7
通讯作者：
Mosher,DF
中科院分区：
文献类型：
--
作者：
Oberley,TD;Murphy-Ullrich,JE;Albrecht,RM;Mosher,DF
原代肾小球细胞放置在含有Way茅斯MB 752/1培养液的化学定义的培养液中，其中添加了胰岛素、转铁蛋白、成纤维细胞生长因子、非必需氨基酸、丙酮酸钠和抗生素，显示出形态上类似于高分化的内脏上皮细胞的细胞迅速生长，随后是低分化的细胞生长；形态证据表明，后者是上皮细胞谱系的前体细胞。尽管通过连续时相显微镜观察从未观察到分化良好的肾小球上皮细胞分裂，但为低分化细胞类型的最佳生长开发了一种化学定义的培养液。这种无血清培养基含Way茅斯的MB 752/1培养基加胰岛素、转铁蛋白、硒和纤维连接蛋白(外加非必需氨基酸、丙酮酸钠和抗生素)。使用这种化学定义的介质，我们比较了二聚体和多聚体纤维连接蛋白(通过将二聚体纤维连接蛋白与3M胍孵育，然后在0.05M环己氨基丙磺酸(CAPS)缓冲液中透析，pH 11，产生高分子量二硫键结合的纤维连接蛋白)对低分化原代肾小球细胞类型黏附和生长的影响。二聚体纤维连接蛋白(FN)在促进肾小球细胞黏附方面的效果是多聚体FN的两倍，尽管这两种形式的FN促进细胞黏附的效果都好于未包被的基质。相反，细胞生长研究表明，多聚体FN是比二聚体FN更有效的生长刺激剂。FN的二聚体和多聚体在体外的不同作用表明，这些分子在体内可能具有不同的功能。
Primary glomerular cells placed in a chemically defined medium containing Waymouth's medium MB 752/1 supplemented with insulin, transferrin, fibroblast growth factor, non-essential amino acids, sodium pyruvate, and antibiotics showed rapid outgrowth of cells which morphologically resembled well differentiated visceral epithelial cells followed by outgrowth of poorly differentiated cells; morphologic evidence suggests these latter cells are precursor cells of the epithelial cell lineage. Whereas the well differentiated glomerular epithelial cells were never observed to divide by sequential phase microscopic observations, a chemically defined medium was developed for optimal growth of the poorly differentiated cell type. This serum-free medium contained Waymouth's medium MB 752/1 supplemented with insulin, transferrin, selenium, and fibronectin (plus non-essential amino acids, sodium pyruvate, and antibiotics). Using this chemically defined medium, we have compared the effects of dimeric and multimeric fibronectin (high molecular weight disulfide-bonded fibronectin produced by incubation of dimeric fibronectin with 3 M guanidine followed by dialysis against 0.05 M cyclohexylaminopropane sulfonic acid (CAPS) buffer, pH 11) on the adhesion and growth of the poorly differentiated primary glomerular cell type. Dimeric fibronectin (FN) was twice as effective as multimeric FN in promoting glomerular cell adhesion, although both forms of FN promoted cell adhesion better than an uncoated substratum. In contrast, cell growth studies demonstrated that multimeric FN was a more potent growth stimulant than dimeric FN. The differential effects of dimeric and multimeric forms of FN in vitro suggests that these molecules may have different functions in vivo.