The Influence Of Chorogenic Acid On Rat Bone Marrow Mesenchymal Stem Cells Culture And Differentiation In Vitro

Background:Studies show that chlorogenic acid is the one of the main active ingredient in Eucommiae ulmoides, with fill the air, strong bones, strong role of will. Objective:research the influence of Chlorogenic acid on rat bone marrow mesenchymal stem cells culture and differentiation in vitro Design, time and place:a randomized controlled animal cell experiments, from March 2010 to September 2010 at the Second Affiliated Hospital of Nanchang University, Central Laboratory of Molecular.MATERIALS:12 clean grade of 2 weeks old SD rats, chlorogenic acid from the Jiangxi Traditional College of Pharmacology Laboratory. Methods:Under sterile conditions, take 2 weeks SD rat femur and tibia bone marrow, taken by density gradient centrifugation and adherent cells were isolated and cultured rat isolated Combination of bone marrow mesenchymal stem cells, containing 10% fetal bovine serum The LG-DMEM culture medium, take the third generation of cells used in the experiment. Take 1mg of chlorogenic acid, dissolved in DMSO in cryopreservation solution, followed by chlorogenic acid diluted with 10-8,10-9,10-10,10-11 g/L of reagents, adding the experimental group of mesenchymal Stem cells. Reagent blank control group and traditional group osteogenic agent group, was observed under inverted microscope every day in the cell growth and morphological changes in cultured 24,48,72 hours a MTT cell proliferation assay, the culture Week 3 was detected by Von Kossa calcium nodules, immunohistochemistry the expression of collagen typeⅡ.Results:12 rats entered the final analysis. MTT method confirmed the experimental group and control group 24 hours after passage, cell proliferation is slow, no significant difference between the two groups (p> 0.05),48小时differences between the two groups (p<0.05),72 hour access to cells in logarithmic growth phase The two groups significantly different (p<0.01).The experimental group were observed bone marrow mesenchymal stem cells in a large number of cells induced to differentiate into osteoblasts, osteogenic agent with the traditional no significant difference, Von Kossa calcium nodules that were identified by traditional bone calcium nodules induced large dose group, Regular shape, color depth, while the experimental group of irregular nodules of calcium, a light color, with the decline in the concentration of the calcium nodules were not evident with traditional osteogenic agent was no significant difference. Immunohistochemistry results showed that both traditional and osteogenic agent group or chlorogenic acid group, not the expression of collagen typeⅡ.Conclusion:chlorogenic acid can promote in vitro bone marrow mesenchymal stem cell proliferation and differentiation into osteoblasts, but its effect and the concentration was some positive correlation may not have the effect of differentiation into Chondrogenic...