Dog and rat kidney proximal tubule cells in culture: responses to parathyroid hormone.
We have described and compared culture systems for proximal tubule cell (PTC) preparations from dog and rat kidney. Cells were prepared from kidney cortex by enzyme digestion and purified on Percoll density gradient. The dog PTC and rat PTC differed in their growth characteristics in culture. Although the dog PTC tended to overgrow the contaminating fibroblastic cells, the rat PTC tended to be overgrown by the latter cells when cultured in medium containing 15% fetal calf serum (FCS). Cultures of rat PTC in serum-free medium or medium containing only 2% FCS yielded only epithelium-like cells exhibiting characteristics of cells that are proximal tubular in origin. These properties include protrusion of microvilli, high alkaline phosphatase activity, ability to transport sugar, and responsiveness to parathyroid hormone (PTH) in terms of cAMP production. The time course and dose-response curves of PTH-stimulated cAMP accumulation were studied in dog and rat PTC. The estimated half-maximal concentrations (Kact) for PTH in dog and rat PTC were 1.2 and 100 nM, respectively. Both values are within the range reported in the literature for the respective renal membrane preparations. In addition to our previously reported data on dog PTC, this study revealed the presence of PTH-inhibitable 25-hydroxyvitamin D3-24-hydroxylase in dog PTC. These two model cell culture systems should prove useful in studying PTH action in renal cells.
Rao
L G
LG
Division of Endocrinology and Metabolism, St. Michael's Hospital, Toronto, Ontario, Canada.
Yau
C H
CH
Kovacs
K
K
D015650:24,25-Dihydroxyvitamin D 3; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002112:Calcifediol / Q000378:metabolism; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000096:biosynthesis; D004100:Dihydroxycholecalciferols / Q000378:metabolism; D004285:Dogs; D005947:Glucose / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D008757:Methylglucosides / Q000378:metabolism; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D010281:Parathyroid Hormone / Q000502:physiology*; D051381:Rats
D004100:Dihydroxycholecalciferols; D008757:Methylglucosides; D010281:Parathyroid Hormone; D015650:24,25-Dihydroxyvitamin D 3; C027020:methylglucoside; D000242:Cyclic AMP; D000469:Alkaline Phosphatase; D005947:Glucose; D002112:Calcifediol