Systematic optimization of the clonal growth of human primary breast carcinoma cells.

摘要: Abstract Human breast carcinomas have been one of the most difficult tumor types to culture in agar-based clonogenic assays. This fact has limited their clinical applicability. We used statistically motivated experimental designs systematically improve clonal enzymatically monodispersed primary human carcinoma cells an anchorage-independent agar system. Based upon initial comparison two basal media, we selected which gave best colony growth and then sought optimize individual additives medium. Hydrocortisone, fetal bovine serum, red blood all improved both plating efficiency median size colonies derived from cells. Next, concentrations these three components were simultaneously idealized using response surface methodology. By methods, it was found that optimal concentration hydrocortisone 0.35 µg/ml, serum 6.5%, 2.1 × 10 7 cells/ml. Using conditions, achieved efficiencies 0.39% 0.19% for with diameters greater than 50 (50 cells) or 70 (130 µm, respectively.