A preparative suspension culture system permitting quantitation of anchorage-independent growth by direct radiolabeling of cellular DNA.
We have developed a hybrid methylcellulose/agar suspension culture system which permits long-term colony formation of transformed mesenchymal cells. In contrast to traditional agar suspensions, our system allows for recovery of cells and direct biochemical analysis of anchorage-independent growth. The ability to readily radiolabel cellular macromolecules in these preparative cultures permits a quantitative and objective analysis of colony formation by incorporation of [3H]thymidine into newly synthesized DNA.
Assoian
R K
RK
Department of Biochemistry and Molecular Biophysics, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Boardman
L A
LA
Drosinos
S
S
D000362:Agar; D002452:Cell Count / Q000379:methods; D002455:Cell Division; D002471:Cell Transformation, Neoplastic / Q000473:pathology*; D002478:Cells, Cultured; D003470:Culture Media; D004261:DNA Replication; D005840:Gentian Violet; D008747:Methylcellulose; D010641:Phenotype; D013535:Suspensions; D013936:Thymidine; D014316:Tritium
D003470:Culture Media; D013535:Suspensions; D014316:Tritium; D000362:Agar; D008747:Methylcellulose; D005840:Gentian Violet; D013936:Thymidine