Research Of Human Marrow Mesenchymal Stem Cell Perfusion Bioreactor Culture And The Biocompatibility Study Of MSC On Scaffolds

AIM: To set up the perfusion bioreactor system and methods for expanding the seed cells in vitro used for tissue engineering, e.g. human mesenchymal stem cell (MSC). To establish a method for evaluating the biocompatibility of biomaterials based on MSC.METHODS: A series of perfusion bioreactor system was designed and assembled according to the principal of animal cell bioreactor culture, and the MSC was performed perfusion culture using this system after the function test and performance test finished by C2C12 cells. The morphology, proliferation, metabolism and osteo-differentiation of the MSC in/post-bioreactor culture were observed and determined, and glucose, lactic acid, LDH and ALP as the metabolism indexes. In biocompatibility study, the morphology of MSC was analysed after directly contacted to the biomaterial with dCMP by SEM in different time points. The concentration of elements in the degradation products of the biomaterials with dCMP or HA were quantitated with ICP and 1C, and the effects of the degradation products on the cell circle of the MSC was evaluated by FACS, on osteo-differentiation of the MSC was assayed quantitatively by ALP and ARS respectively.RESULTS: The structure of perfusion bioreactor is rational. The results of the perfusion bioreactor culture indicated that the bioreactor system is safe and feasible to culture cells, because C2C12 cultured in it proliferated significantly better than in traditional T-flask culture while its morphology wasn't affected. Also, the morphology, metabolism, proliferation and differentiation of MSC cultured in it weren't affected. The efficiency of media was improved. In biocompatibility study, dCMP enhanced the proliferation of MSC, but didn't interfere the osteo-differentiation of MSC and its mineralization. And, HA inhibited the proliferation process of MSC and the mineralization of osteo-differentiated MSC, while it did not interfere with the osteo-differentiation process of MSC.CONCLUSION: MSC could be expanded in this perfusion bioreactor system and methods in vitro. A method for evaluating the biocompatibility of biomaterials based on MSC was estabolished. dCMP had a better cytocompatibiltiy with MSC than HA, which might allow for its use as skeleton scaffolds.