1. Separation of renal medullary cells: isolation of cells from the thick ascending limb of Henle's loop
- Author
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Jill Eveloff, Rolf K. H. Kinne, and Winfried Haase
- Subjects
Arginine ,Cell Survival ,Hydrolases ,Population ,Ficoll ,Cell Separation ,Biology ,Ouabain ,chemistry.chemical_compound ,Adenosine Triphosphate ,Oxygen Consumption ,Loop of Henle ,medicine ,Animals ,education ,Kidney ,education.field_of_study ,Cell Biology ,Articles ,Molecular biology ,Organoids ,medicine.anatomical_structure ,Kidney Tubules ,Biochemistry ,chemistry ,Alkaline phosphatase ,Trypan blue ,Rabbits ,Sodium-Potassium-Exchanging ATPase ,medicine.drug ,Adenylyl Cyclases - Abstract
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.
- Published
- 1980