Multiple Hematopoietic Growth Factors Secretion By Rats Bone Mesenchymal Stem Cells Stimulated With Astragaloside Iv

Objective:To explore a practical method of isolation and culture of the rat MSCs in vitro and to observe their living characteristics and to offer an experimental foundation for their further differentiation and actual application. To study the effect of astragaloside IV on the expression of multiple cytokines by bone mesenchymal stem cells(MSCs)in rats.Methods:MSCs were isolated from adalt Wistar rats and cultured by using the method of adhesive screening and clone, then detected the biological activity: the cell surface antigen of MSCs was detected with immunocytochemistry.Proliferation of MSCs stimulated with astragaloside IV was ascertained by the MTT method. Expression of hematopoietic growth-factors was ascertained using RT-PCR in MSCs with astragaloside IV or not.Results:The isolated MSCs of the mononuclear cells of marrow, mostly subcultured cells adhered the wall in 12~24h, presented the colony type growth, exhibiting a large expansive potential and high suitability, a typical fibroblastlike morphology, and 92.3%expressing CD44 by immunocytochemistry, only 6.9%expressing CD45. MSCs were effectively isolated and purified in vitro, and expressed of many hematopoietic growth-factors except IL-3, such as stem cell factor(SCF), thrombopoietin(TPO), granulocyte macrophage colony stimulating factor(GM-CSF)and transforming growth factor(TGF-β1). Astragaloside IV stimulation promoted MSCs proliferation, and 200mg/L astragaloside IV treatment produced a peak effect 72h after culture. The expression of SCF stimulated with astragaloside IV increased significantly compared with that in MSCs without astragaloside IV stimulation.Conclusions:MSCs could be effectively isolated and purified by using the method of adhesive cultiration and clone, meanwhile, they can express many hematopoietic growth-factors. Astragaloside IV may promote MSCs proliferation and increase SCF secretion in vitro.