Effects Of Proliferation And Osteogenic Potentiality In Rat Bone Marrow Mesenchymal Stem Cell By Using BrdU Labeling

Objective :To observe the effects of proliferation and osteogenic potentiality of rat bone marrow mesenchymal stem cells (MSCs) by using BrdU labeling.Methods :Rat marrow derived-MSCs were isolated and adherent cultured . The ability of monoclone and proliferation were detected after labeling with 0.2% BrdU. Meanwhile the MSCs were induced to osteoblast by osteogenic liquid for 3 weeks. Osteogenic potentiality was observed by alizarin bordeaux , fluorescence immunohistochemistry and Real Time PCR.Results :The proliferation and monoclone of MSCs were decreased obviously after BrdU labeling. The number and area of clones in labled group were less than they were in unlabeled group (P<0.05). The growing curve showed that the proliferation of the labeling group was significantly lower than it was in unlabled in 6 d, 8 d, 12 d and 14 d, and the peak was lower than the unlabled one. As the osteogenic potentiality, two groups cells were expressed osteocalcin and collagen and were appearenced calcium nodules. However, the RT-PCR reported that there was no significant difference in the expression of osteocalcin gene between the two groups (p>0.05).Conclusions :The proliferation and monoclone were inhibited by labeled BrdU, but the ability of the osteogenic was not be decreased. Therefore, BrdU labeling can be applied widely on the field of MSCs as seed cells for the investigation of stem cells.