The Capacity To Multilineage Differentiation Of Rat Bone Marrow Mesenchymal Stem Cells And Analyses Of Time Sequence Change Of Differentiation To Related Genes

Objective: Aim to build a simple and effective method of primary cell culture?induced differentiation and staining of bone marrow mesenchymal stem cells(BMSC) and observe the biological characteristics and potential capacity of BMSC to differentiate to osteoblasts and adipocytes and detect the related gene expression changes in the process of cell differentiation in order to prepare for the experiment about difference of gene function.Methods: BMSC was separated by whole bone marrow culture and induced by differentiation induced solution. We discussed the methods of separation?culture and the growth law and the conditions to differentiate to osteoblasts and adipocytes by observing the form change of BMSC and staining identification of osteoblasts and adipocytes during differentiation. Determining the expression of ALP?OCN?Runx2?PPAR??FABP4 by RT-PCR.Results:(1) We successfully separated ?cultured BMSC and draw the growth curve of BMSC of passage 4;(2)Identified the of osteogenic potential by alizarin red staining;(3) Identified the of osteogenic potential by Oil red O staining. Conclusion The purity of BMSC separated by whole bone marrow culture can reached a certain level after subcultured for 4 times and the BMSC can differentiate to osteoblasts and adipocytes under special conditions which can be identified by certain stainings;(4) Determining the expression of related genes by RT-PCR.Conclusions: The purity of BMSC separated by whole bone marrow culture can reached a certain level after subcultured for 4 times and the BMSC can differentiate to osteoblasts and adipocytes under special conditions which can be identified by certain stainings. Along with the prolonged the expression of related genes(ALP?OCN?Runx2?PPAR??FABP4) has changed,so it has the own function of adjustion.