A microplate assay for quantitation of anchorage-independent growth of transformed cells

摘要: Abstract We developed a 96-well microplate assay to quantitate anchorage-independent growth of transformed cells. Wells tissue culture microtiter plates are coated with poly(2-hydroxyethyl methacrylate) (poly(HEMA)) prevent cell attachment, and cells suspended in liquid media seeded into the treated plates. Cell is assessed by tetrazolium dye reduction or counting [3H]thymidine incorporation DNA. There appeared be close correlation between poly(HEMA)-coated colony formation soft agar, i.e., fibroblasts various oncogenes proliferated plates, whereas their normal counterparts did not. Transformed on nonadhesive surface were round formed multicellular spheroids which resembled colonies agar. Cells carrying either temperature-sensitive v-src inducible v-Ki-ras oncogene only when they displayed phenotype. This method simple, quick, quantitative, economical many cases might more practical than conventional agar for measurement growth.