Influence of hyaluronan on proliferation and differentiation of BMP-2 transfected BMSCs of goats

Objective To evaluate the effect of hyaluronan(HA)on proliferation and differentiation of the adenovirus mediated bone morphogenetic protein-2 gene(Adv-BMP-2)transfected goats bone marrow derived mesenchymal stem cells(BMSCs)in vitro.Methods The BMSCs were separated from the marrow of the goats and expanded,and then been divided into the follow five groups:Adv-BMP-2 transfected BMSCs+HA(group 1),Adv-BMP-2 transfected BMSCs(group 2),Untransfected BMSCs+HA(group 3),Untransfected BMSCs as the contrast group(group 4),Adv-β-gal transfected BMSCs(group 5).The proliferation of the BMSCs was evaluated by count the average amount of BMSCs,plot the growth curve of the BMSCs and flow cytometry,and the differentiation of the cells was assessed by the alkaline phosphatase(ALP)assay and RT-PCR for the mRNA of collagen-Ⅰ(Col-Ⅰ),osteonectin(ON) and bone sialoprotein(BSP).Results BMSCs proliferation: There were no significant different between these groups after three days culture,but the group 1,3 were significant higher than the other groups in the 7th day.The ALP assay: After 3 days culture,the ALP activity was decreased in group3,whereas increased in group 1 and 2;but after 7 days culture,all the group 1,2,3 were significant higher than the group 4,5.The mRNA expressions of Col-Ⅰ,ON and BSP:In the 3 rd and 7th day after cultured,all the mRNA expression was enhanced in the group 1,2,3.Conclusions HA may accelerate BMSCs proliferation,increase ALP activity and the gene expression of Col-Ⅰ,ON,and BSP when BMSCs are cultivated with HA in vitro.