Susceptibility of primary cultures of proximal tubular and distal tubular cells from rat kidney to chemically induced toxicity.
Isolated proximal tubular (PT) and distal tubular (DT) cells from rat kidney were cultured for up to 9 days under serum-free, hormonally-defined conditions on 35-mm polystyrene culture dishes. Several hormonal and growth factor supplements were assessed for their ability to promote growth (increased protein and DNA content) and stability of differentiated phenotype (high activities of gamma-glutamyltransferase and alkaline phosphatase as brush-border membrane markers in PT cells; maintenance of high activities of glutamate dehydrogenase as a mitochondrial marker in both PT and DT cells; maintenance of low and high activities of lactate dehydrogenase in PT and DT cells, respectively; expression of cytokeratins). Basal supplemented media (DMEM/F12, 1:1 v/v) contained insulin, hydrocortisone, epidermal growth factor, sodium selenite and transferrin as supplements. Additionally, triiodothyronine selectively promoted growth and stability of differentiated phenotype in PT cells and thyrocalcitonin selectively promoted growth and stability of differentiated phenotype in DT cells. On Day 3 of primary culture, PT and DT cells were incubated for up to 8 h with either tert-butyl hydroperoxide (tBH; 0.5-10 mM), methyl vinyl ketone (MVK; 1-10 mM), or p-aminophenol (PAP; 1-10 mM) and cellular injury, as assessed by cellular release of lactate dehydrogenase, was determined. DT cells were significantly more susceptible to injury from both tBH and MVK, but the two cell populations were equally susceptible to injury from PAP, which is the same susceptibility pattern seen in freshly isolated cells. These results suggest that primary cultures of rat renal PT and DT cells reflect similar biochemical properties as freshly isolated cells and are, therefore, useful models for study of chemically induced injury.
Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
L H
LH
Lash
J J
JJ
Tokarz
D M
DM
Pegouske
D000469:Alkaline Phosphatase / Q000378:metabolism; D000627:Aminophenols / Q000633:toxicity*; D000818:Animals; D002074:Butanones / Q000633:toxicity*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004247:DNA / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D007686:Kidney Tubules, Distal / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D008858:Microscopy, Phase-Contrast; D008871:Microvilli / Q000187:drug effects / Q000201:enzymology; D000073718:Pancreatitis-Associated Proteins; D010545:Peroxides / Q000633:toxicity*; D011506:Proteins / Q000378:metabolism; D051381:Rats; D011916:Rats, Inbred F344; D017382:Reactive Oxygen Species / Q000633:toxicity*; D005723:gamma-Glutamyltransferase / Q000378:metabolism; D020122:tert-Butylhydroperoxide
D000627:Aminophenols; D002074:Butanones; D016895:Culture Media, Serum-Free; D000073718:Pancreatitis-Associated Proteins; D010545:Peroxides; D011506:Proteins; C000612141:REG3A protein, human; D017382:Reactive Oxygen Species; D004247:DNA; D020122:tert-Butylhydroperoxide; C057920:3-buten-2-one; D007770:L-Lactate Dehydrogenase; D005969:Glutamate Dehydrogenase; D005723:gamma-Glutamyltransferase; D000469:Alkaline Phosphatase; C026729:4-aminophenol