Primary cultures of rabbit renal proximal tubule cells: I. Growth and biochemical characteristics.
Before the usefulness of a new in vitro model can be ascertained, the model must be properly defined and characterized. This study presents the growth rate and biochemical characteristics of rabbit renal proximal tubule cells in primary culture over a 2-wk culture period. When grown in a hormonally defined, antibiotic-free medium these cells form confluent monolayer cultures within 7 d after plating. Multicellular dome formation, an indicator of transepithelial solute transport, was expressed after confluent cultures were formed. The activity of the cytosolic enzyme, lactate dehydrogenase, and the lysosomal enzyme, N-acetyl-glucosaminidase, increased 14- and 2-fold during the first 8 d of culture, respectively. In contrast, the activity of a brush border enzyme, alkaline phosphatase, decreased 85% within the first 8 d of culture. Release of these enzyme markers into the culture medium, which are routinely used to measure cytotoxicity, stabilized after 8 d in culture. The ratio of cellular protein to DNA changed according to the state of cellular growth. Values rose from 0.035 mg protein/micrograms DNA in preconfluent cultures to 0.059 mg protein/micrograms DNA in confluent cultures. These results document the characteristics of a primary proximal tubule cell culture system for future studies in in vitro toxicology.
Aleo
M D
MD
Department of Pharmacology & Therapeutics, State University of New York, Buffalo 14214.
Taub
M L
ML
Nickerson
P A
PA
Kostyniak
P J
PJ
D000118:Acetylglucosaminidase / Q000032:analysis / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000032:analysis / Q000378:metabolism; D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D003470:Culture Media; D004247:DNA / Q000032:analysis / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000032:analysis / Q000166:cytology* / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000378:metabolism; D008297:Male; D011817:Rabbits
D003470:Culture Media; D004247:DNA; D007770:L-Lactate Dehydrogenase; D000469:Alkaline Phosphatase; D000118:Acetylglucosaminidase