Development and characterization of rabbit proximal tubular epithelial cell lines.
Development and characterization of rabbit proximal tubular epithelial cell lines.
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兔近端肾小管上皮细胞系的发育和表征。
DOI：
10.1038/ki.1992.397
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发表时间：
1992
影响因子：
19.6
通讯作者：
Jacobberger,JW
中科院分区：
文献类型：
--
作者：
Romero,MF;Douglas,JG;Eckert,RL;Hopfer,U;Jacobberger,JW
兔近端肾小管上皮细胞系的建立及特性。我们分离了兔肾近端小管上皮细胞系。选择的基础是它们在多孔支架上形成融合单层的能力，以及维持受体介导的信号转导和离子传输的能力，这是近端小管的特征。该分离方法包括几个步骤：(1)显微切割皮质浅层近端小管，并将其培养在基质涂层的多孔支架上，直到细胞形成融合的单层；(2)选择具有激素调节的近端小管典型离子转运的原代培养物，并与辐射后的成纤维细胞共培养；(3)将传代多次后存活的上皮细胞在多孔性基质上扩增和传代。以这种方式建立的大多数细胞系是通过与辐射后的成纤维细胞共培养而获得的，该细胞产生编码SV40大T抗原和G418抗性的重组逆转录病毒。然而，SV40和T抗原的表达对永生化并不是必不可少的，因为在分离的细胞系中既没有检测到T抗原，也没有检测到G418抗性，并且与不产生3T3的细胞共同培养也得到了类似的结果。其中一个细胞系(VEPT)已经在形态、生化和离子转运特性方面进行了一些详细的表征。这条线形成融合的单层，顶端有微绒毛，紧密连接，基侧质膜卷曲。一旦融合，单分子层在培养数周内保持25至32ms/cm2的电导，并在含葡萄糖的培养液中具有根瘤菌素敏感的短路电流(ISC)，这表明顶端Na+-葡萄糖共转运。VEPT细胞还保留了血管紧张素II(Ang II)的受体和信号机制。顶端和基端的血管紧张素II和5，6-环氧二十碳三烯酸(5，6-EET)以与原代培养相似的方式调节ISC。这些细胞系与原代培养细胞共享细胞角蛋白K8、K10/K11和K19的表达(命名法[21])。它们还保留了几种受体和信号转导机制。例如，Ang II、花生四烯酸、缓激肽、5，6-EET、甲状旁腺激素(残基1至34)和嘌呤核苷酸增加胞浆钙离子，PTH提高cAMP水平，Ang II增强近端小管特异性花生四烯酸代谢。
Development and characterization of rabbit proximal tubular epithelial cell lines. We have isolated rabbit kidney proximal tubular epithelial cell lines. The selection was based on their ability to form confluent monolayers on porous supports and to maintain receptor-mediated signal transduction and ion transport, characteristic of the proximal tubule. The isolation method consisted of several steps: (1) superficial cortical proximal tubule segments were microdissected and cultured on a matrix-coated porous support until cells formed a confluent monolayer; (2) primary cultures showing hormone-regulated ion transport typical for the proximal tubule were selected and co-cultured with irradiated fibroblasts; and (3) the epithelial cells surviving after several passages were expanded and passaged on porous substrates. Most of the cell lines developed in this manner were obtained by co-culture with irradiated fibroblasts producing a recombinant retrovirus encoding SV40 large T antigen and G418 resistance. However, SV40 T antigen expression was not essential for immortalization, since neither T antigen nor G418 resistance was detected in the isolated cell lines and co-culture with non-producing 3T3 cells gave similar results. One cell line (vEPT) has been characterized in some detail with respect to morphological, biochemical, and ion transport properties. This line forms confluent monolayers with apical microvilli, tight junctions, and convolutions of the basolateral plasma membrane. Once confluent, monolayers maintain conductances of 25 to 32 mS/cm2for several weeks in culture and possess phlorizin-sensitive short-circuit current (Isc) in glucose containing media, indicative of apical Na+-glucose co-transport. vEPT cells also retain receptor and signaling mechanisms for angiotensin II (Ang II). Apical and basal Ang II and 5,6-epoxyeicosatrienoic acid (5,6-EET) modulate the Isc in a manner similar to primary cultures. The cell lines share with primary cultures expression of the cytokeratins K8, K10/K11, and K19 (“nomenclature” [21]). They also retain several receptor and signal transduction mechanisms. For example, Ang II, arachidonate, bradykinin, 5,6-EET, parathyroid hormone (residues 1 through 34), and purine nucleotides increase cytosolic Ca2+, PTH elevates cAMP levels, and Ang II enhances proximal tubule-specific arachidonic acid metabolism.
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兔肾小管上皮细胞磷脂酶 C 活化与前列腺素 E2 和环磷酸腺苷生成之间的关系。
DOI：
10.1172/jci113376
发表时间：
1988
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Welsh,C;Dubyak,G;Douglas,JG
通讯作者：
Douglas,JG
建立源自近端小管 S2 段的肾细胞系。
DOI：
10.1159/000173395
发表时间：
1991
期刊：
Renal physiology and biochemistry
影响因子：
--
作者：
C. Fauth;D. Chabardès;M. Allaz;M. García;B. Rossier;F. Roch;M. Claire
通讯作者：
M. Claire
兔肾皮质来源的 SV40 转化管状细胞系近端和远端细胞功能的维持
影响因子：
5.6
作者：
A. Vandewalle;B. Lelongt;M. Géniteau;B. Baudouin;M. Antoine;S. Estrade;F. Châtelet;P. Verroust;R. Cassingena;Pierre M. Ronco
通讯作者：
Pierre M. Ronco
根根苷对体外葡萄糖和水肠道转运的作用
DOI：
10.1113/jphysiol.1958.sp006109
发表时间：
1958
期刊：
The Journal of Physiology
影响因子：
--
作者：
B. Parsons;D. Smyth;C. B. Taylor
通讯作者：
C. B. Taylor
影响因子：
64.8
作者：
D. Hand;P. A. Sanford;D. Smyth
通讯作者：
D. Smyth