In Vitro Bromodeoxyuridine Labeling Method of Human Carcinomas under Various Conditions

Abstract

The proliferative activity of tumor cells in human gastric and colonic adenocarcinoma was measured by means of in vitro incorporation of the thymidine analogue, 5-bromo-2'-deoxyuridine (BrdU), into the newly-synthesized DNA of fresh tumors. The incorporation was optimized by incubation for 2 hours at 3-4 atm. of mixed gas (95% 02 + 5% C02) with 100 flM BrdU admixed in the culture media. Fresh specimens up to 2 hours after resection retained their ability to incorporation, but their S-phase fraction gradually decreased thereafter. The corporation rate was preserved better by refrigeration than by storing at room temperature. With the advantage of its safety and reliability in evaluation of the proliferative activity of human malignancy, in vitro BrdU labeling technique could be recommended as a routine procedure to assess the prognosis of the patient.