A Further Study of the Mode of Action of Methylcholanthrene on Normal Tissue Cultures

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Abstract
The action of purified methylcholanthrene has been studied on 3 series of fibroblasts from adult C3H strain mice. These cell strains were grown in Carrel flasks in chicken plasma, horse serum and chick embryo extract, and were subjected to 0.01 mg. per cc. of culture fluid, for 114 days, 230 days, and 145 days, respectively, after which they were removed to fresh culture fluid in clean culture flasks, and carried for various times up to about 260 days without further addition of carcinogen. Controls were run, with no methylcholanthrene, for each series. The cells were shielded from white light and subjected only to light of wave length longer than 480 m[mu], in order to eliminate any photosensitizing action of the methylcholanthrene. The initial effect of the carcinogen was toxic and retardative for growth. This toxicity seemed possibly less pronounced at later periods. Even 260 days after removal from the carcinogen, the cells had not reassumed their normal morphology and physiology. The cultures previously treated with the carcinogen showed central necrosis of the culture unless the cultures were transferred to a fresh flask every 14 days, the controls could be carried uninterruptedly without change of flask for from 60 to 140 days. While the controls showed the fibroblasts as normal, laterally discrete cells, spindle shaped, with slender, thread like terminal processes, the cells which had once been treated with the carcinogen showed a striking degree of lateral adhesion. The cells were apparently permanently changed from the action of the carcinogen, and this change was such as to render the cells very closely similar to cells from tumors which had arisen in rats from methylcholanthrene inj.

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