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Growth and characterization of human skin epithelial cell cultures

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Summary

In 129 of 140 attempts, human skin cells were successfully cultured on the dermal collagen bed of sterile, dead pigskin. Diploid epithelial cells grew selectively on the collagen bed; fibroblasts grew on the glass surfaces of the culture dishes. The cultures could be subdivided physically up to six times at a 1:2 split ratio, but at least 24 to 48 cell generations were produced over the months the cells could be carried. Much of the cell multiplication resulted in maturation into distinct basal, squamous, granular, and keratinized cell layers. The cultured cells were considered epithelial because of their shape, possession of intercellular bridges, desmosomes and tonofibrils, and because they formed maturating epithelium in vitro and upon transplantation back to the original human donor. As the cells grew they digested the pigskin collagen, thus producing clear zones that could be used to monitor and quantitate cell growth. Multiplication of epithelial cells, rather than migration, was indicated by mitotic figures in colchicine-treated cultures and by DNA synthesis.

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Authors and Affiliations

  1. The Children’s Hospital of Akron, 44308, Ohio

    Aaron E. Freeman, Howard J. Igel, Brenda J. Herrman & Karen L. Kleinfeld

Authors
  1. Aaron E. Freeman
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  2. Howard J. Igel
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  3. Brenda J. Herrman
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  4. Karen L. Kleinfeld
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Additional information

Expert technical assistance was provided by Nancy Allen (cell culture); William Towler (electron microscopy); James Malone, Nona Scaife, and Joy M. Nicolet (cytogenetics); R. Thomas Campbell and Dorothy Sarver (photography); and V. L. Angerstein, Susan Ekker, and Arnater Yarbrough (histology).

This work was supported by The United Fund Cancer Society of Summit County, the Greater Cleveland Associated Foundation (grant no. 3G3490X1), the National Institute of General Medical Services (grant no. 1 R01 GM 21929-01), and the Charles E. Merrill Trust.

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Freeman, A.E., Igel, H.J., Herrman, B.J. et al. Growth and characterization of human skin epithelial cell cultures. In Vitro Cell.Dev.Biol.-Plant 12, 352–362 (1976). https://doi.org/10.1007/BF02796313

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