Experimental Stduy On The Effect Of Labeling SD Rat Bone Marrow Mesenchymal Stem Cells With Edu

Objective:Lableing the SD rat bone marrow mesenchymal stem cells with a different concentration of Edu.Exploring the optimal concentration and time of bone marrow mesenchymal stem cells from Edu lableing.Methods:(1) Separate and purify BMSC via marrow adherence, observe cellular morphology, detect cell cycle surface marker via flow cytometer(FCM) and then induction identify the osteogenic and adipogenic differentiation.(2) Mark the BMSC by Edu at the concentration of0,10,20and50umol/L and at the time of12,24,48and72hours, measure the labeling rate via FCM. Pick out the optimal labeling concentration at12,24,48and72hours, measure the positive rate of Edu marked cells at each concentration and point-in-time, observe the marked cellular morphology, draw the growth curve of cell and manage the experiment of activity detection, multiplication and induced differentiation after cell marking.Results:(1) Identification of BMSC:morphologic characters:adherent cells appeared24-48hours after inoculation, most of them are round, polygon or short shuttle-like, cellular morphology was uniform in the third generation, grew adhering to the wall in single layer, short shuttle-like shape and spiral pattern, flow cytometry detection of surface marker CD90,CD29,CD45and CD11b showed94.3%,99.4%,3.2%and5.7%respectively, dyeing with oil red after induced14days by adipogenesis inducer, hyacinth lipid droplet was observed under microscope, dyeing with alizarin red after induced14days by osteogenesis inducer, hyacinth calcium nodule was observed under microscope, blueviolet calcium nodule was observed under microscope by alkaline phosphatase staining.(2) Labeling rate in each concentration group went up with extension of labeling time (P<0.05), labeling time in each time group went up with elevation of labeling concentration, but the difference was not significant(P>0.05). Conclusion:The labeling method with Edu is sample but efficacious, and the optimal group is labeling with the concentaation of lOumol/L and the time with48h.