Characterization and Spontaneous Neoplastic Transformation of Mouse Embryo Cells Isolated and Continuously Cultured In Vitro in Chemically Defined Medium NCTC 135

Publisher Website
Google Scholar
Abstract
A strain of fibroblast-like cells has been isolated from C3H mouse embryos and grown in chemically defined medium NCTC 135. These cells were isolated by mechanical disaggregation of the source tissue and have never been exposed to protein supplements, enzymes, or hormones. Colonies of epithelium-like cells were observed after 249 days' growth in vitro and these cells appeared to have a lethal effect on the fibroblast-like cells when the two cell types came into contact. After the epithelium-like cells were eliminated by mechanical means, the strain of fibroblast-like cells was finally developed. By the time the chromosomal analyses were initiated on the cells of this strain, the mode was hypotetraploid. The cell strain does not yet exhibit a stabilized mode. These cells can be grown in some of the less complex formulations of the NCTC media containing vitamin B12 and i-inositol which are required for continuous growth of this strain. They can also be grown continuously in medium NCTC 135 produced by at least one commercial source. The strain can also be successfully frozen and stored in liquid nitrogen with dimethylsulfoxide as the protective agent. Malignant transformation in vitro, as evidenced by tumor production, was first noted from intraocular implants of cells in culture 411 days. The tumors were transplantable and were composed of fusiform cells. Histologically the tumors resembled the fibrosarcomas commonly derived from tissue-cultured cells.