Problems of in vitro culture of human mammary tumor cells

Abstract

Various techniques were used to obtain cell cultures from benign and malignant tumors removed from human breasts. Cell suspensions were prepared by trypsinizing finely minced tissue or by the 'spilling' technique of Lasfargues and Ozzello, and T flasks or T flasks coated with collagen were inoculated. Trypsinized tissue yielded predominantly fibroblastic cultures, whereas those obtained by the 'spilling' technique were rich in epithelial cells. The mixed cell populations were separated into pure suspensions by density gradient centrifugation. Most epithelial cell cultures grew best when they were collected by the 'spilling' technique and inoculated into collagen coated T flasks with Eagle's medium enriched with 30% fetal calf serum. However, tears developed in the collagen coat with subsequent peeling which led to the loss of many epithelial cell colonies. Primary epithelial cells did not remain viable for prolonged periods, and attempts to obtain subcultures were unsuccessful. In this paper the work of several investigators is reviewed and their methods of growing breast tumor cells are discussed.