A homogeneous population of single cells from the thick ascending limb of Henle's loop (TALH) has been isolated from the rabbit kidney medulla. A total medullary cell suspension was prepared by a series of collagenase, hyaluronidase, and trypsin digestions and separated on a Ficoll gradient (2.6-30.7% wt/wt). Morphologically, the cells isolated from the TALH were homogeneous and showed polarity within their plasma membrane structure, with a few blunt microvilli on their apical surface and deep infoldings of the basal-lateral membrane. Biochemically, the TALH cells were highly enriched in calcitonin-sensitive adenylate cyclase and Na, K-ATPase. Alkaline phosphatase and arginine vasopressin-sensitive adenylate cyclase, highly concentrated in proximal tubule and collecting duct, were present only in low concentrations in the TALH cells. Additionally, furosemide, a diuretic inhibiting sodium chloride transport in the TALH in vivo, inhibited oxygen consumption of the TALH cells in a dose-dependent manner. The TALH cells were viable, as judged by morphological appearance, trypan blue exclusion, the response of oxygen consumption to 2,4-dinitrophenol, succinate and ouabain, and the cellular Na, K and ATP levels.
Article| December 01 1980
Separation of renal medullary cells: isolation of cells from the thick ascending limb of Henle's loop.
Online ISSN: 1540-8140
Print ISSN: 0021-9525
J Cell Biol (1980) 87 (3): 672–681.
J Eveloff, W Haase, R Kinne; Separation of renal medullary cells: isolation of cells from the thick ascending limb of Henle's loop.. J Cell Biol 1 December 1980; 87 (3): 672–681. doi: https://doi.org/10.1083/jcb.87.3.672
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