INVITRO BRDURD INCORPORATION OF COLORECTAL TUMOR-TISSUE

This study describes a novel in vitro method for the incorporation of the thymidine analogue, bromodeoxyuridine (BrdUrd), in fresh colorecta l tumour tissue. Disaggregation by pronase, collagenase and DNAse resu lted in high cell yields of viable single cell suspensions, representa tive of the original tumour, which could be infiltrated with BrdUrd. A modified ELISA identified optimal incubation times and BrdUrd concent rations. This technique has been used in preliminary studies to invest igate two important areas intrinsic in the analysis of BrdUrd colorect al cell proliferation data: 1 to determine the effects of the individu al constituents of the cell culture media, in particular glutamine, on BrdUrd incorporation in suspensions of colorectal cells and 2 to exam ine the denaturation step. This method will have wide applicability in investigations of cell proliferation status in both normal and diseas ed tissue.