The Influence Of Proliferation And Ostogenic Differentiation Of1,25(OH)2VD3on HUCMSCs In Vitro

Background and ObjectiveMesenchymal stem cells (MSCs) has strong self-renewal capacity and multi-differrentiation potential. As the seed cells, MSCs has broad application prospects in the field of cell therapy, tissue engineering and regenerative medicine. It can provide new treatment options for the major diseases of intractable diseases, such as peripheral nerve injury, spinal cord injury, bone defects, orthopedic. Compared to hES, hBMSCs and ADSC, as the ideal seed cells with good biological characteristics, hUCMSCs was easy to separated and has low immunogenicity and strong differentiation capacity. In vitro, culture conditions was the governor decided to induce stem cell-specific differentiation to a certain extent. Appropriate inducer such as vitamin C, glycerophosphate, dexamethasone, and some growth factors can be effectively induce MSCs to differentiate into osteoblasts. Traditional osteogenic fluid was complex and toxic, to establish a new efficient and safe osteogenic methord has been the focus of the study of people. This study was To establish the separation and expansion method of human umbilical cord derived mesenchymal stem cells(hUCMSCs) and explore the influence of1,25(OH)2VD3on hUCMSCs in vitro, and to find a new method inducing them to differentiate into osteoblasts for Bone tissue engineering. MethodshUCMSCs were cultured from wharton’s jelly of human umbilical cord in the condition of sterilitas. hUCMSCs were cultured from adherent tissue pieces and detected by immunohistochemistry, surface antigens of hUCMSCs were detected by immunohistochemistry; Taking the third generation with good growth state hUCMSCs make intervene experiment. Set10-7mol/L、10-9mol/L、10-11mol/L3kind of concentration of1,25(OH)2VD3as the experimental group, and a control group (DMEM medium). The morphologic change of UCMSCs was observed by inverted microscope. Observe the cell proliferation by the method of MTT. Detect alkaline phosphatase assay and type I collagen assay from immunohistochemistry. And the calcium tubercle formation would be examined after21days. The results were statistically analyzed.ResulthUCMSCs were cultured from adherent tissue pieces. Under microscope, an initial hBMSCs was round, stretched out processes gradually, formation cells of long strips similar to fibroblasts.1W later swirled growth amplificated rapidly. hUCMSCs strongly positive for CD4、CD105and negative for CD34、CD45consistent with the hUCMSCs characteristics. Under different concentration of1,25(OH)2VD3hUCMSCs proliferation were promoted within7days but were suppressed beyond7days. The high doses group (10-7mol/L group) obvious inhibitted the stem cell proliferation. Different concentrations of1,25(OH)2VD3and days have interactive effect (P<0.05). The differences between different groups with ALP and type I collagen assay has statistical difference(P<0.05). The secretion of ALP and type I collagen assay of10-7mol/L concentration group was higher than others. Typical mineralization nedus was found in intervene groups.ConclusionThe hUCMSCs can be successfully cultured from the adherent tissue pieces.1,25(OH)2VD3can effectively induce hUCMSCs to differentiate into osteoblasts in vitro in specific conditions.1,25(OH)2VD3can be used as cell activity factors and have a broad prospect in osteonecrosis, Bone tissue engineering and bone defect.