Use of monoclonal antibodies to culture rat proximal tubule cells.
Use of monoclonal antibodies to culture rat proximal tubule cells.
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使用单克隆抗体培养大鼠近曲小管细胞。
DOI：
10.1152/ajpcell.1986.251.5.c780
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Seifter,JL
中科院分区：
文献类型：
--
作者：
Stanton,RC;Mendrick,DL;Rennke,HG;Seifter,JL
目前的肾细胞培养技术受到细胞产量低或细胞类型异质性的限制。我们已经使用单克隆抗体的微绒毛膜蛋白分离和培养的近曲小管细胞的纯人口。通过相差显微镜、碱性磷酸酶、丁酸酯酶和γ-谷氨酰转移酶的酶组织化学、电子显微镜和与各种近端小管细胞单克隆抗体的特异性反应，将细胞表征为近端小管细胞。在2-7天的生长产生的细胞数量高达1,000倍以上，通过单小管显微切割获得。观察到圆顶形成，表明液体转运完整。此外，Na+-H+交换和Na+依赖的D-己糖转运，已知的近端小管的运输过程，分别通过单细胞和甲基-α-D-吡喃葡萄糖苷摄取的显微荧光法证明。我们的研究结果表明，大量的同质的，培养的大鼠近端小管细胞，保持在体内近端小管细胞的特性，可以获得使用单克隆抗体技术的分离。
Current renal cell culture techniques are limited by either a low yield of cells or by heterogeneity of cell types. We have used monoclonal antibodies to microvillus membrane proteins to isolate and culture a pure population of proximal tubule cells. The cells were characterized as proximal tubule cells by phase microscopy, enzyme histochemistry for alkaline phosphatase, butyrate esterase, and gamma-glutamyltransferase, electron microscopy, and specific reactivity with a variety of monoclonal antibodies for proximal tubule cells. Growth over 2-7 days yielded cell numbers up to 1,000-fold greater than obtained by single tubule microdissection. Dome formation was observed, suggesting intact fluid transport. In addition, Na+-H+ exchange and Na+-dependent D-hexose transport, known transport processes of the proximal tubule, were demonstrated by microfluorimetry of single cells and methyl-alpha-D-glucopyranoside uptake, respectively. Our results indicate that large numbers of homogeneous, cultured rat proximal tubule cells that maintain characteristics of in vivo proximal tubule cells can be obtained using a monoclonal antibody technique of isolation.