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Variation in Capacity for Anchorage-independent Growth among Agar-derived Clones of Spontaneously Transformed BALB/3T3 Cells¹

Department of Molecular Biology, University of California, Berkeley, California 94720

A subline of cloned spontaneously transformed BALB/3T3 cells had a colony-forming efficiency (CFE) in agar of 5 to 20%. Individual agar colonies isolated and reseeded into agar were not significantly more efficient at initiating colonies than the original unselected subline. Four successive cycles of agar growth and selection also failed to increase the mean CFE in agar. Randomly selected clones isolated on a plastic surface all had the capacity to grow in agar. These results suggest that the failure of the majority of the cells to grow in agar is not the result of an intrinsic or heritable inability to do so. The ability to initiate a colony in agar seems to vary phenotypically from cell to cell. In contrast, agar colonies isolated from some tumor cell lines (originating from related spontaneously transformed 3T3 cells) and reseeded in agar had a higher CFE than the unselected tumor cell lines. In one case, this increased CFE in agar was lost when the cells were passaged on plastic without further selection for agar growth. Thus, expression of the anchorage-independent phenotype may vary, even among related cloned populations of transformed cells.

¹ This research was supported by USPHS Grant CA 15744 from the National Cancer Institute and United States Department of Energy, Office of Environment Contract AT03-79EV10277.

² To whom requests for reprints should be addressed.

Received 4/16/84. Accepted 8/24/84.