The development of serum-free hormone-supplemented media for primary kidney cultures and their use in examining renal functions

Abstract

We have demonstrated the primary kidney cultures and defined medium can be used for several purposes. First, primary kidney cultures can be examined in defined medium to evaluate the physiological significance of studies made with established kidney cell lines. Secondly, new kidney epithelial cell culture systems can be developed, using primary cultures and defined medium. The studies presented here have demonstrated several possible approaches towards these ends. First, a hormone-supplemented serum-free medium (Medium K-1) developed for an established dog kidney epithelial cell line (MDCK) was used to grow primary cultures of baby mouse kidney epithelial cells without fibroblast overgrowth. The primary cultures of baby mouse cells not only grew in response to each of the five supplements in Medium K-1, but also possessed several of the differentiated kidney functions observed in the MDCK cell line. Secondly, defined media were developed for primary cultures of rabbit kidney epithelial cells directly. The rabbit cells were shown to require a different set of supplements than MDCK cells to attain optimal growth. Furthermore primary cultures derived from purified rabbit proximal tubules required a different set of supplements than primary cultures derived from unpurified rabbit kidney tissue. These new primary culture systems should prove invaluable in examining renal transport functions in vitro.