Human glomerular visceral epithelial cells synthesize a basal lamina collagen in vitro.
Human glomerular visceral epithelial cells synthesize a basal lamina collagen in vitro.
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人肾小球内脏上皮细胞在体外合成基底层胶原。
DOI：
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发表时间：
1979
影响因子：
11.1
通讯作者：
G. Striker
中科院分区：
文献类型：
--
作者：
P. Killen;G. Striker
用纯化的细菌胶原酶消化分离的人肾小球，产生上皮细胞。这些细胞生长至饱和密度并且没有变成多层。通过相差显微镜和电子显微镜观察其形态外观以及 C3b 表面受体的存在，将它们鉴定为内脏肾小球上皮细胞。未观察到因子 VIII 抗原和 Fc 受体。肾小球上皮细胞合成了一种胶原蛋白，其抗原性与人肾小球基底层相似。使用针对非胶原肾小球基底层抗原的亲和纯化抗体从内脏上皮细胞培养基中沉淀蛋白质，产生单一胶原酶不稳定蛋白质，通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳，在还原剂存在下以 168,000 的表观 Mr 迁移。羟脯氨酸异构体分析得出3-羟脯氨酸与总羟脯氨酸的比率为0.17。胃蛋白酶消化产生二硫键多聚体，通过还原，其迁移的表观 Mr 为 148,000。这些数据表明，人肾小球内脏上皮细胞可以在体外分离和增殖，并且它们合成与体内发现的胶原相似的胶原。
Isolated human glomeruli were digested with purified bacterial collagenase yielding epithelial cells. These cells grew to saturation density and did not become multi-layered. They were identified as visceral glomerular epithelial cells by their morphologic appearance by phase and electron microscopy and by the presence of surface receptors for C3b. Neither Factor VIII antigen nor Fc receptors were observed. The glomerular epithelial cells synthesized a collagenous protein that was antigenically similar to human glomerular basal lamina. Proteins precipitated from visceral epithelial cell medium with affinity purified antibody against noncollagenous glomerular basal lamina antigens yielded a single collagenase labile protein that by sodium dodecyl sulfate/polyacrylamide gel electrophoresis migrated with an apparent Mr of 168,000 in the presence of reducing agents. Analysis of hydroxyproline isomers yielded a ratio of 3-hydroxyproline to total hydroxyproline of 0.17. Pepsin digestion yielded a disulfide-bonded multimer which, with reduction, migrated with an apparent Mr of 148,000. These data demonstrate that human glomerular visceral epithelial cells can be isolated and propagated in vitro and that they synthesize a collagen similar to that found in vivo.