BMP-2,BGP mRNA Expression Quantity and ALP Active Change to Rat Bone Marrow Mesenchymal Stem Cell in Vitro Induction Osteoblasts Influence

Objective To establish a system of isolating and cultivation of bone mesenchymal stem cells(BMSCs) from the bone marrow in vitro and to study the ability of osteoinductive induced by BMSCs.The effect of the expression of BMP-2,BGPmRNA and the changes of ALP activity on proliferation and differentiation of osteoblast were also studied.Methords Rat bone marrow stromal cells(BMSCs) were isolated by the methods of density centrifugation and adhesive culture.Cell morphology was observed,BMP were induced into ostoblast by the mechanism of osteoinductive.The ALP activity and the exp ression of BMP-2,BGP mRNA were detected quantitatively by observation of cell morphology,histochemical staining,enzyme immunoassay and RT-PCR on the 7d and 14d.Results High pure BMSCs was obtained by the methods of density centrifugation combined with adhesive culture.Both ALP activity and mineraliztion node staining were positive in bmscs after osteogenic differentiation 7d and 14d.osteogenic differentiation group:the activity of ALP(82±1.0 U/L),the expression of BMP-2(6.04±0.02),BGP mRNA(3.21±0.02) on the 14 d were significantly higher than that on the 7d.Uninduced group:the activity of ALP was low.No expression of BMP-2,BGP mRNA,no formation of the mineralized nodules either.The results showed that:along with prolongation of the culture time,the expression of BMP-2 became higher.The ALP activity and the expression of BGP mRNA were higher also.Cells showed changes in morphology differentiating into osteoblasts.All these indicate:there was a relation between the formationg of osteoblast.Conclusion BMSCs could have been induced to osteoblast successfully and a system of isolating and cultivation in vitro was established.It is suggested that osteoinductive has a high relativity with the increase of the ALP activity and the higher expression of the BMP-2,BGPmRNA.