Research Of Rat Marrow Mesenchymal Stem Cell Culture And In Vitro Cytotoxicity Study Of MSC On Novel Materials

Objective: The study explore the separation and the characteristics of mesenchymal stem cells (MSCs) derived from rat bone marrow and messure the cytotoxicity of a novel, biodegradable aliphatic/aromatic copolyesters by MSCs-co-material culturing method.Methods: The MSCs were isolated from rat bone marrow by density gradient centrifuge and adhesive property; immunohistochemistry was applied to detect the expression of CD29,CD34 and CD71,and growth curve was drawn to compare the separation and proliferation of different passage. In cytotoxicity study, the cell viability, growth, mitochondrial metabolic activity, membrane integrity and morphology were determined and observed with four types of assays[Neutral Red, amido black, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetranolium bromide , acridine orange/ethidium bromide]and by scanning electron microscopy using extract and direct contact methods.Results: The cells separated by density gradient centrifuge and adhesive property owned the characteristics of MSCs, and was verified by the positive result of CD29,CD71 and negative result of CD34. The biomaterials tested did not induce significant cytotoxic effects and exhibited normal metabolic function and cell growth in vitro. Conclusions: The MSCs can be successfully separated by density gradient centrifuge and adhesive property. The three copolyesters of different constitunent ratio of mass is absence of significant cytotoxicity. In cytotoxicity assays, Neutral Red uptake assay and amido black staining method are sensitive and reliable, inexpensive, easily handly, so that, they may be one of the major methods of cytotoxicity assay or bio- compatibility assay in common laboratory.