An Experiment Study About Isolation,culture Of Mouse Adipose-derived Stem Cells In Vitro And Factors' Secretion Related To The Hematopoietic Regulation Of It And After It's Induced To Osteoblasts

Objective: To investigate the isolation,culture of mouse adipose-derived stem cells (ADSCs) and factors'secretion related to the hematopoietic regulation of it and it's induced to osteoblasts at molecule level in vitro. Methods: ADSCs were isolated by digesting adipose tissue with collagenase typeⅠ, and be expanded to three passages after primary culture in a control medium. Then the cells were incubated in adipogenic medium and osteogenic medium for 2～3weeks. These factors'secretion forβ-catenin,SDF-1,N-cadherin,SCF,MIP-1αand Jag1, GM-CSF were confirmed by real-time quantitative PCR. While identification about surface markers of adipose-derived stem cells such as CD29,CD34,CD106,CD49d by flow cytometry. Results: ADSCs grew as adherent cells and presented fibroblast-like in the control medium in vitro, and could stably proliferate and be passed. The cells'surface antigen phenotype were analyzed by flow cytometry, and th expression of CD29 was strongly positive, CD34 was lower, the expression of CD106 and CD49d was negative. ADSCs incubated in the adipogenic medium were stained positively for oil-red O and incubated in the osteogenic medium were stained positively for alzarin red staining and von Kossa staining. There were no statistical differences of expression of factors'genes between ADSCs and after it's induced to osteoblasts through detection analysis of real-time quantitative PCR (P>0.05). The expression ofβ-catenin was the highest. Followed by SDF-1 and N-cadherin. The expression of the other factors such as SCF,MIP-1α,Jag1.,GM-CSF were all lower. Conclusion: In this study, the cells from adipose tissue were ADSCs. The expression of factors'genes related to the hematopoietic regulation has different degrees in ADSCs and after it's induced to osteoblasts, but has no statistical differences between them.