Ruhfus B, Bauernschmitt H G, Kinne R K
Max-Planck-Institut für molekulare Physiologie, Abteilung Epithelphysiologie, Dortmund, Germany.
In Vitro Cell Dev Biol Anim. 1998 Mar;34(3):227-31. doi: 10.1007/s11626-998-0128-4.
A primary culture from rat renal IMCD cells was established to investigate the permeability characteristics of the luminal and contraluminal plasma membranes of the papillary collecting duct in vitro. Freshly isolated IMCD cells were grown on filters in a special "epithelial cell" medium. Confluency was proved with an epithelial volt/ohm meter. After 7 d of culture the transepithelial resistance reached more than 1000 omega x cm2. A polarization of the cells with regard to a basolateral localization of a lactate efflux system, and an L-alanine transport system was achieved. The hypotonicity-activated release systems for the organic osmolytes sorbitol and betaine were also located basolaterally, whereas taurine, glycerophosphorylcholine, and myo-inositol left the cells at both cell poles but with different capacity. Morphological observations revealed also that the monolayer was well differentiated. Thus, a model of a renal collecting duct epithelium was established which can be used to analyze polarized and differentiated transport processes across the epithelial cells and their plasma membranes.
建立了大鼠肾内髓集合管（IMCD）细胞的原代培养体系，以体外研究肾乳头集合管腔面膜和对侧腔面膜的通透性特征。将新鲜分离的IMCD细胞接种于滤器上，置于特殊的“上皮细胞”培养基中培养。用上皮伏特/欧姆计检测细胞融合情况。培养7天后，跨上皮电阻超过1000Ω·cm²。细胞实现了乳酸外排系统和L-丙氨酸转运系统在基底外侧的极化分布。有机渗透剂山梨醇和甜菜碱的低渗激活释放系统也位于基底外侧，而牛磺酸、甘油磷酸胆碱和肌醇在细胞两极均可离开细胞，但转运能力不同。形态学观察还显示单层细胞分化良好。因此，建立了一种肾集合管上皮模型，可用于分析跨上皮细胞及其质膜的极化和分化转运过程。
