FAST AND SENSITIVE METHOD FOR SIMULTANEOUS MEASUREMENT OF CELL-PROLIFERATION RATE AND DRUG-SENSITIVITY IN RAT CEREBRAL-CORTEX

A proliferation assay based on the production of mini-units of tissue was adopted and modified for the simultaneous determination of cell pr oliferation rate and the effect of genistein in rat cerebral cortex. M ini-units of tissue were produced from rat cerebral cortex immediately after killing the animal and incubated with culture medium containing H-3-methyl-thymidine during 90 min. The proliferation rate was assess ed by measurement of H-3-methyl-thymidine incorporation into trichloro acetic acid insoluble material/mg of protein/min. The mini-unit method preserves the neural-cell topological relation existing in vivo and, in addition, has several additional advantages: (1) the short incubati on time required limits the metabolic changes, (2) the sensitivity to drugs can be assessed simultaneously with the cell proliferation rate, (3) the complete procedure can be performed within 4-6 h, and (4) man y experiments can be performed with the tissue from one animal. Genist ein in doses from 10 to 100 mu M inhibited cell proliferation in a con centration-dependent manner. The percentage of inhibition was highest in young animals and decreased with increasing age. This method is a p owerful tool for the study of drugs with short-time onset mechanisms o f action and can be useful for the screening of new drugs. (C) 1998 Ac ademic Press.