Isolation, culture and characterization of human renal proximal tubule and collecting duct cells.
Isolation, culture and characterization of human renal proximal tubule and collecting duct cells.
复制标题
人肾近曲小管和集合管细胞的分离、培养和表征。
DOI：
10.1159/000020633
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Trifillis,AL
中科院分区：
文献类型：
--
作者：
Trifillis,AL
人类肾单位在细胞类型方面的复杂性和异质性使得明确定义的肾细胞体外系统对于研究肾毒性中涉及的发病机制是有价值的。在我们的实验室中，肾近端小管细胞（PTC）和集合管细胞（CDC）已从尸体肾（死后时间<24 h）中分离、培养和表征，用于治疗剂和细菌诱导的肾细胞毒性的研究。以106个/ml接种的PTC在7天内形成融合单层。使用组织化学标记物来确定细胞培养物的来源。细胞对因子VIII呈阴性，对细胞角蛋白和γ-谷氨酰转移酶（GGT）呈阳性，并结合翅豌豆凝集素。从肾乳头分离的CDC在接种后14天内形成单层。CDC均为VIII因子和GGT阴性，细胞角蛋白和结合花生凝集素阳性。PTC和CDC分离株和培养物表现出典型的上皮细胞超微结构：细胞连接，中间丝，微绒毛，和许多线粒体。形态学和组织化学证据证实，PTC和CDC可以分离和培养用于体外研究。
The complexity and heterogeneity of the human nephron with regard to cell types make well-defined in vitro systems of renal cells valuable for studies of the pathogenetic mechanisms involved in nephrotoxicity. In our laboratory renal proximal tubule cells (PTC) and collecting duct cells (CDC) have been isolated, cultured and characterized from cadaver kidneys (postmortem time <24 h) for use in studies of renal cytotoxicity induced by therapeutics and bacteria. PTC seeded at 106cells/ml formed confluent monolayers within 7 days. Histochemical markers were used to determine the origin of the cell cultures. Cells were negative for factor VIII, positive for cytokeratin and γ-glutamyltransferase (GGT), and bound winged pea lectin. CDC, isolated from the renal papillae, formed monolayers within 14 days of seeding. CDC were negative for factor VIII and GGT, positive for cytokeratin and bound peanut lectin. PTC and CDC isolates and cultures exhibited typical epithelial cell ultrastructure: cell junctions, intermediate filaments, microvilli, and numerous mitochondria. The morphological and histochemical evidence confirms that PTC and CDC can be isolated and cultured for use in in vitro studies.