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Mouse Epidermal Cells and Carcinogenesis I. Isolation of Skin Constituents¹

( McArdle Memorial Laboratory, The Medical School, University of Wisconsin, Madison 6, Wisconsin)

A method has been developed for the enzymatic dissection of mouse skin into its constituents and their further dispersion into individual cells. The skins from the backs of plucked, depilated mice are incubated in a solution of Eagle's minimal medium with hyaluronidase. The loosened dermis and hair follicles are scraped away. Incubation is then continued with elastase, which loosens the epidermis and permits epidermal sheets to be scraped away. The epidermis can then be dispersed into individual cell suspensions mechanically or with trypsin. The basal cells can be obtained in almost pure form by gradient centrifugation in Eagle's medium plus albumin. Serial specimens of cells can be obtained from living mice by gentle squeezing following intradermal injection of trypsin.

¹ This work was supported by Grant # CA-07175 of the National Cancer Institute, National Institutes of Health, USPHS. A preliminary account of this work appeared in Proc. Am. Assoc. Cancer Research 4:23 (1963).

² American Cancer Society Professor of Oncology.

Received 8/17/64.