An established cell line from mouse kidney medullary thick ascending limb. I. Cell culture techniques, morphology,...

The effective use of cultured cells as model systems for investigating the differentiation and regulation of transport processes in renal tubular epithelial cells depends on the availability of functional long-term cell lines derived from specific nephron segments. Conventional culture procedures that treat cells as proliferating microorganisms possess several inherent limitations that could contribute to phenotypic instability and limited proliferative capacity in vitro. In this study, culture techniques were adopted that avoid exposure of cells to proteolytic enzymes, maintain intercellular contacts, and allow cells to remain continually adherent to a collagen gel substratum. This methodology resulted in the development of a continuous epithelial cell line from identified, microdissected segments of the mouse kidney medullary thick ascending limb.