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Isolation of Variant Cells from SV40-transformed Human Diploid Fibroblasts¹

University of Vermont, Burlington, Vermont 05401 [N. R., W. I. S.], and Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104 [L. D.]

Variant cell lines that are sensitive to density-dependent inhibition of growth have been isolated from three of four SV40-transformed human fibroblast cell lines. Variants were isolated by plating the transformed cells at low density, by treating them with 5-fluorodeoxyuridine, or by growing them on glutaraldehyde-fixed monolayers of normal cells. The variant cell lines, isolated at a frequency of about 2% of all cells forming colonies after treatment, were initially recognized by colonial morphology, and the variant phenotypes were confirmed, after subculturing, by saturation-density determinations. The variant cell lines reach saturation densities that are 40% or less than those of the parent cell lines, and plate in soft agar medium at reduced efficiency, compared with the parent cells. They retain SV40 T antigen. The modal chromosome numbers of two of the variant cell lines were increased, compared with those of the parent cell lines; two other variants were indistinguishable in chromosome number from the parent cells. Stability of these properties over a 6-month period was demonstrated with two of the variants.

¹ This investigation was supported in part by USPHS Grants CA 08936, CA 10815, and CA 12056 from the National Cancer Institute, HD 06323 from the National Institute of Child Health and Human Development, and RR 05540 from the Division of Research Resources; and by the Commonwealth of Pennsylvania.

² Present address: Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Mass. 02139.

Received 2/ 3/75. Accepted 4/22/75.