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Growth factor requirements of organogenesis in serum-free metanephric organ culture

  • Rapid Communications in Cell Biology
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Summary

In order to define humoral growth factors which may regulate mammalian renal development, the growth requirements of fetal metanephric organogenesis were studied in serum-free murine organ culture. Metanephric growth, determined by cell proliferation and protein content, and metanephric differentiation, determined morphometrically as epithelial glomerular formation, were compared and contrasted following 144 hours of organ culture incubation in basal medium, basal medium supplemented with 10% fetal bovine serum, and basal medium supplemented with various combinations of growth factors. The basal medium was composed of equal volumes of Dulbecco's modified Eagle's medium and Hams' F-12 medium. Five humoral growth factors were studied in the following concentrations: selenium, 6.8×10−9 M; insulin, 8.3×10−7 M; triiodothyronine, 2×10−9 M; transferrin, 6.2×10−8 M; and prostaglandin E1, 7.1×10−8 M. Results showed that transferrin and prostaglandin E1 were necessary for optimal growth in the system and that prostaglandin E1 was necessary for maximal metanephric differentiation. Such data provide guidelines for the creation of serum-free medium for future fetal renal cell and tissue culture systems, and provide insight into the factors which may regulate normal and abnormal renal embryogenesis and the reparative processes of renal hyperplasia and hypertrophy which follow renal injury.

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Authors and Affiliations

  1. Department of Pediatrics, Division of Nephrology, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania

    Ellis D. Avner, William E. Sweeney Jr., Nicholas P. Piesco & Demetrius Ellis

  2. University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania

    Ellis D. Avner, William E. Sweeney Jr., Nicholas P. Piesco & Demetrius Ellis

Authors
  1. Ellis D. Avner
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  2. William E. Sweeney Jr.
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  3. Nicholas P. Piesco
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  4. Demetrius Ellis
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Additional information

A preliminary report of this work was presented at the Ninth International Congress of Nephrology, Los Angeles, June 1984. These studies were supported in part by Basil O'Connor Starter Research Grant 5-349 from the March of Dimes Birth Defects Foundation and New Investigator Research Award I-R23-AM34891-01 from the National Institute of Arthritis, Diabetes, and Digestive and Kidney Diseases of the National Institutes of Health (Both to Dr. Avner).

Editor's Statement The determination of effects of extracellular components on the introduction and maintenance of differentiation is an area for which serum-free culture techniques are particularly suited. The approaches described in this report utilize morphometric techniques to quantitate differentiation in serum-free fetal organ culture in addition to standard methodologies for assessing growth. The purely epithelial nature of the cultures used in these studies also provides some interesting advantages in the design of experiments aimed, at determining the importance of cell-cell interactions at various stages in the differentiative process. David W. Barnes

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Avner, E.D., Sweeney, W.E., Piesco, N.P. et al. Growth factor requirements of organogenesis in serum-free metanephric organ culture. In Vitro Cell Dev Biol 21, 297–304 (1985). https://doi.org/10.1007/BF02620946

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  • DOI: https://doi.org/10.1007/BF02620946

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