Fifty-one new (CCRF) cell lines have been established in vitro from the primary cultures of 142 specimens of normal and neoplastic tissues from human and animal sources during a 2½-year study. Twenty-four of these cell lines were established from human, and 27 cell lines from animal, sources. The cell lines derived from animal sources include P-1534 and L1210 leukemia, an Ehrlich ascites tumor, and a spontaneous leiomyosarcoma of Syrian hamsters. There was no difference in the frequency of successful isolation of cell lines from human and animal tissue, but such frequency was greater from non-neoplastic tissue, irrespective of species, in this series of experiments.

The amino acid requirements of the CCRF cell lines in general are similar to those described for other cell lines propagated in these basal media. The kind of serum used as supplement was of critical importance for the serial propagation of the majority of cell lines of animal origin. Whole calf serum, or dialyzed calf serum supplemented with m-inositol, permitted the isolation and serial propagation of animal cell lines which failed to grow in the same basal medium supplemented with horse serum. This difference between calf and horse serum was not associated with the attachment of cells to glass, since the cells in aliquots of the same specimen became attached to the surface of the flask when either serum was used but did not survive and grow in media supplemented with horse serum.

The data presented indicate that, with minor modifications, the basal media described by Eagle et al. are suitable for the direct isolation of mammalian cells from a variety of biopsy specimens. However, certain kinds of human neoplasia consistently have failed to grow, suggesting that the present media may not be optimal for all cells.

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These studies were supported in part by Research Grants CY-2782 and CY-3335 from the National Cancer Institute of the National Institutes of Health, United States Public Health Service.

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