Summary
A new mouse metanephric organ culture system has been developed to study mammalian renal development. The system permits in vitro organotypic differentiation in a serum-free, hormone supplemented medium consisting of Dulbecco’s minimal essential medium (MEM) and Ham’s F12 medium supplemented with insulin, 5 µg/ml; PGE1, 25 ng/ml; T3, 3.2 pg/ml; hydrocortisone, 5 µg/ml; and transferrin, 5 µg/ml. In this system, metanephric development continues morphologically beyond the S-shaped tubule stage. A well differentiated proximal tubule forms with a well defined brush border, specialized intercellular connections, and an apical endocytic network. In addition, a unique devascularized glomerulus, with highly differentiated podocytes surrounding areas of basement membrane, forms entirely from epithelial elements.
The present organ culture model goes beyond the limitations of previously described systems in that it does not require separation of nephrogenic blastema from ureteric bud, nor require animal serum or nonspecific tissue extracts for metanephric development. The model is thus suited for morphological, biochemical, and endocrinological study of normal and abnormal renal organogenesis.
Similar content being viewed by others
References
Grobstein, C. Inductive epithelio-mesenchymal interaction in cultured organ rudiments of the mouse. Science 118: 52–55; 1953.
Saxén, L.; Koskimics, O.; Lahti, A.; MicHinen, H.; Rapola, J.; Wartiovaara, J. Differentiation of kidney mesenchyme in an experimental model system. Adv. Morphogen. 7: 251–291; 1968.
Gossens, C. L.; Unsworth, B. R. Evidence for a two-step mechanism operating duringin vitro mouse kidney tubulogenesis. J. Embryol. Exp. Morphol. 28: 615–631; 1972.
Emura, M.; Tanaka, T. Development of endothelia and erythroid cells in the mouse metaneophrogenic mesenchyme in culture with the fetal liver. Dev. Growth Diff. 14: 237–246; 1972.
Borghese, E.; Dani, A. M. Observations on the differentiationin vitro of mouse metanephros. Wilhelm Roux’ Arch. Entwickl.-Mech. Org. 164: 83–96; 1969.
Taub, M.; Chuman, L.; Saier, M.; Sato, G. Growth of Madin-Darby canine kidney epithelial cell (MDCK) line in hormone-supplemented, serum-free medium. Proc. Natl. Acad. Sci. USA 76: 3338–3342; 1979.
Taub, M.; Sato, G. Growth of functional primary cultures of kidney epithelial cells in defined medium. J. Cell. Physiol. 105: 369–378; 1980.
Gruneberg, H. The development of some external features in mouse embryos. J. Hered. 34: 88–92; 1943.
Vainio, T.; Jainchill, J.; Clement, K.; Saxén, L. Studies on kidney tubulogenesis VI. Survival and nucleic acid metabolism of differentiating mouse metanephrogenic mesenchymein vitro. J. Cell Comp. Physiol. 66: 311–318; 1965.
Larsson, L. The ultrastructure of the developing proximal tubule in the rat kidney. J. Ultrastruct. Res. 51: 119–139; 1975.
Bernstein, J. Morphologic development of the metanephric tubule. Proc. 7th Int. Congr. Nephrol. (Montreal) 1978: 249–254.
Avner, E. D.; Villee, D.; Grupe, W. E. Ontogeny of proximal tubular antigen (FX1A) in metanephric organ culture (abstr.). Kidney Int. 19: 179; 1981.
Pegg, D.; Bernstein, J.; Hook, J. B. Biochemical and ultrastructural correlates of substrate stimulation of renal organic anion transport. Proc. Soc. Exp. Biol. Med. 151: 720–725; 1976.
Reeves, W.; Caulfield, J. P.; Farquhar, M. G. Differentiation of epithelial foot processes and filtration slits: Sequential appearance of occluding junctions, epithelial polyanion, and slit membranes in developing glomeruli. Lab. Invest. 39: 90–100; 1978.
Bernstein, J.; Cheng, F.; Roszka, J. Glomerular differentiation in metanephric culture. Lab. Invest. 45: 183–190; 1981.
Potter, E. Development of the human glomerulus. Arch. Pathol. 80: 241–255; 1965.
Kazimierczak, J. Development of the renal corpuscle and the juxtaglomerular apparatus: A light and electron microscopic study. Acta Pathol. Microbiol. Scan. [A] (Suppl. 218): 1–64; 1971.
Grobstein, C. Inductive interaction in the development of mouse metanephros. J. Exp. Zool. 130: 319–355; 1955.
Grobstein, C. Transfilter induction of tubules in mouse metanephrogenic mesenchyme. Exp. Cell. Res. 10: 424–440; 1956.
Gluecksohn-Waelsh, S.; Rondon Rota, T. Development in organ tissue culture of kidney rudiments from mutant mouse embryos. Dev. Biol. 7: 432–444; 1963.
Rapola, J.; Niemi, M. Studies in kidney tubulogenesis. Cytochemical localization of phosphatase and dehydrogenase activities during the formation of tubulesin vitro. Z. Anat. Entwickl. Gesch. 124: 309–320; 1965.
Wartiovaara, J. Cell contacts in relation to cytodifferentiation in metanephrogenic mesenchymein vitro. Ann. Med. Exp. Fenn. 44: 469–503; 1966.
Brown, A. L. An analysis of the developing metanephros in mouse embryos with abnormal kidneys. Am. J. Anat. 47: 117–171; 1931.
Ludwig, E. Embryologische beobachtungen an den harnorganen der maus und des goldhamsters. Acta Anat. (Basel) 29: 1–19; 1957.
Norgaard, J. O. R. Retraction of epithelial cell foot processes during culture of isolated glomeruli. Lab. Invest. 38: 320–329; 1978.
Scheinman, J. I.; Fish, A. J.; Kim, Y.; Michael, A. F. C3b receptors on human glomeruliin vitro: Loss in culture. Am. J. Pathol. 92: 147–154; 1978.
Oberly, T. D.; Burkholder, P. M.; Barber, T. A.; Hwang, C. C. Cytochemical characterization of cultured adult guinea pig glomerular cells. Invest. Cell. Pathol. 2: 27–43; 1979.
Oberly, T. D.; Murphy-Ullrich, J. E.; VicMuth, J. The effect of fetal calf serum on the biology of cultured primary glomerular cells. Diagn. Histopath. 4: 117–128; 1981.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Avner, E.D., Ellis, D., Temple, T. et al. Metanephric development in serum-free organ culture. In Vitro Cell.Dev.Biol. -Plant 18, 675–682 (1982). https://doi.org/10.1007/BF02796422
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02796422