SD Rat Bone Marrow Mesenchymal Stem Cells-derived Microvesicle’s Effect On Chondrocyte Proliferation And Phenotype

Objective(1)Isolate and culture SD rat bone marrow mesenchymal stem cells(bone marrowmesenchymal stem cells, BMSCs) by the whole bone marrow adherence method, andidentify the surface marker molecules and differentiation potential of BMSCs;(2)To explore ways of extraction and identification of SD rat bone marrowmesenchymal stem cells-derived microvesicles.(3)To investigate the effect of BMSCs-MVs on SD rats articular chondrocyte’sproliferation and phenotype by co-culture of SD rat BMSCs-MVs and articularchondrocyte in vitro.Methods(1)Isolate and culture SD rat BMSCs, and identify BMSCs by methods of inducing thedifferentiation of stem cells and flow cytometry technology(FCM).(2)Extract microvesicles(MVs) from SD rat BMSCs by ultracentrifugation and identifytheir shape by electron microscopy;(3)Isolate and culture the articular chondrocyte of 2-month-old SD rat;(4)BMSCs-MVs and the third generation chondrocytes were co-cultured, and test theproliferation of the chondrocytes by Ed U and CCK-8 kit;RT-PCR method was usedto detect the changes of synthesis and metabolic activity of co-culturedchondrocytes.Results(1)It was found that BMSCs can be induced to differentiate into chondrocytes and fatcells.FCM analysis showed the expression rate of CD29、CD90 of BMSCs were98.7% and 99.2%, while the rates of CD45、CD11b were and 3%, 0.2%;(2)The activity of chondrocyte’s proliferation was detected by Ed U, CCK-8.Comparingwith control group,The hyperplasia of co-cultured chondrocyte with BMSCs-MVsis obvious(P<0.01),and the higher of the BMSCs-MVs dose,the stronger effect ofthe promoting proliferation(P<0.01).(3)RT-PCR analysis find that comparing with the control group,the expression ofaggrecan(Acan), type II collagen in the experimental group were enhanced, while theexpression of type X collagen, MMP-3 and IL-1βdecrease.Conclusions(1)In vitro, SD rat BMSCs-MVs can promote the proliferation of chondrocytes;(2)In vitro, SD rats BMSCs-MVs can enhance the expression of the Acan and type IIcollagen of the articular chondrocytes, and inhibit the synthesis of the Type Xcollagen 、MMP-3 and IL-1β.It indicated that BMSCs-MVs might be beneficial tothe maintenance of normal phenotype of chondrocytes.