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Established Lines of SV40-transformed Human Amnion Cells¹

Sloan-Kettering Institute for Cancer Research, New York, New York 10021 and Sloan-Kettering Division, Graduate School of Medical Sciences, Cornell University, New York, New York 10021 [J.F.]

Five of 200 cultures, representing 22 strains of SV40-transformed human amnion cells in "crisis," gave rise to established cell lines. In contrast to the frequent crisisrecovery reported for SV40-transformed fibroblasts, only one amnion cell line (A244) was recovered after routine serial propagation of the transformed cells. One line was derived from a strain [H (as)] treated with SV40 antiserum, and three lines originated from replicate cultures of a mycoplasma- and SV40-infected strain [E (Mel)], from which mycoplasma was eliminated. All lines were recovered between the third and sixth month of crisis. Only one of the lines (A244) ceased producing virus, at 115 days after recovery. As reported for nonvirus-producing established lines of SV40-transformed fibroblasts, heterokaryon formation between A244 cells and CV-1 cells resulted in the production of SV40. The cells were susceptible to infection with either SV40 or virus DNA. Comparison of the five lines showed that higher chromosome numbers (80 to 100) were correlated with an increase in polycentrics and new chromosome varieties, a decrease in acentric fragments and minute chromosomes, a shorter population doubling time, and epithelial-like morphology. The lines with the lowest chromosome numbers (50 to 60) were more fibroblastic.

¹ This investigation was supported in part by National Cancer Institute Research Grant CA-08748.

² Recipient of a research fellowship grant from the Westchester Division, American Cancer Society. Present address: Biology Department, Pennsylvania State University, University Park, Pa. 19602.

Received 11/10/69. Accepted 1/29/70.