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Explantation and Subculture of Epithelial Cells from Human Uterine Ectocervix¹

Cancer Research Center [E. H. V., J. G. N., A. M. C., J. S. P.], and the Departments of Obstetrics and Gynecology [L. A. W.], Pediatrics [A. M. C.], Bacteriology and Immunology [J. S. P.], and Medicine [J. S. P.], University of North Carolina, Chapel Hill, North Carolina 27514

Human ectocervical explant cultures were grown in medium with D-valine substituted for L-valine. Pure epithelial cell monolayers were obtained with both dialyzed and undialyzed fetal calf serum. Epithelial cell explant colonies grown in D-valine medium supplemented with undialyzed serum could routinely be subcultured if plated at a density of 1.5 x 10⁴ cells/sq cm or higher. Such cultures could be passaged at least three times and could yield up to 21 population doublings per culture lifetime. Contaminating fibroblastic colonies were never detected in these cultures, which were free of surface-associated fibronectin as revealed by immunofluorescent tests. Both primary and passaged epithelial colonies retained many characteristic morphological features of ectocervical epithelium when examined by light and electron microscopy. Such cultures may be of use in investigating the action of viral and chemical carcinogenic agents upon epithelial cells in vitro.

¹ Supported by NIH Grants CA16086 (University of North Carolina, Cancer Research Center Core Support) and CA19014 (DNA Virus Genomes, Oncogenesis and Latency).

² Recipient of Research Training Fellowship awarded by the International Agency for Research on Cancer. On leave of absence from the Department of Obstetrics and Gynecology, Helsinki University Central Hospital, Helsinki, Finland.

³ To whom requests for reprints should be addressed.

Received 9/18/79. Accepted 11/14/79.