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Experimental Study on Xtracting Solution from Eucommia Ulmoides Oliv Induces Differentiation of Goat Bone Marrow Mesenchymal Stem Cells into Osteoblasts and Inhibits the Differentiation into Adipocytes

Author: ChenWeiCai
Tutor: LuoJun
School: Nanchang University
Course: Surgery
Keywords: Leaf extract MSCs Osteogenic differentiation Cbfa 1
CLC: R68
Type: Master's thesis
Year: 2009
Downloads: 95
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Abstract

Background and purpose: mesenchymal stem cells in the bone marrow (bone marrow mesenehymal stem cells, MSCs) are non-hematopoietic stem cells in the bone marrow, has the ability of self-replication and differentiation potential. Under different conditions can be induced to differentiate into multiple cell lines, including osteoblasts, fibroblasts, chondrocytes, fat cells, muscle cells, endothelial cells and nerve cells. Traditional inducers has simply induced osteogenic or adipogenic differentiation, more limited application. Leaf extract as an inducer Pharmacodynamics improve bone density and inhibit bone resorption, the regulation of bone metabolism, promote bone cell proliferation and alkaline phosphatase secretion while inhibiting adipocyte differentiation. This study investigated the leaf extract induced sheep MSCs to osteoblast differentiation and demonstrate its ability to inhibit the role of MSCs into fat differentiation. Content and methods of research: 1. Using whole bone marrow in vitro isolation and culture of sheep bone marrow mesenchymal stem cells, conventional methods were subcultured MSCs identification, flow cytometry CD29, CD44 and CD105. 2 leaf extract on the proliferation and osteogenic differentiation of MSCs. Detection of ALP, calcium nodules staining to observe promote osteogenic differentiation effect. Eucommia leaf extract inhibits adipogenic differentiation of MSCs role. Oil red-O staining observed inhibition of adipocyte differentiation effects. Eucommia leaf extract of cells MSCs osteogenic differentiation the Cbfa lmRNA expression was detected from the cellular level, automatically gel imaging analyzer analysis observe the effect of promoting osteogenic differentiation. Results: 1 whole bone marrow culture method (ie adherent method) separation of the purified bone marrow mesenchymal stem cells, the medium was changed and trypsin digestion and passage gradually purified, simple and practical method. Compared with the blank control group, the alkaline phosphatase synthesis into the cells of the bone induction group increased formation of mineralized nodules, and traditional osteogenic inducer leaf extract group ,10-4 ,10-5g / mL Eucommia biloba extract group osteoinductive better than traditional osteogenic induction agent group, 10-6g / mL Eucommia leaf extract group. 3 Oil Red O staining results show that adding the leaf extract adipogenic group, the number of lipid droplets was significantly less than traditional adipogenic agent group and the control group, 10-4,10-5 g / mL Eucommia biloba extract group effect. 4. RT-PCR osteogenic induction group could efficient expansion of 171bp of Cbfa 1mRNA gene transcription fragments and 10-4,10-5 g / mL concentration of leaf extract into bone cells of Cbfa 1mRNA in expression level differences no significant (P gt; 0.05). Conclusion: leaf extract can induce isolated and purified culture in vitro of sheep bone marrow mesenchymal stem cells differentiation and proliferation of osteoblasts direction while inhibiting adipocyte differentiation, two-way adjustment. Leaf extract 10-4,10-5 g / mL concentration osteogenesis and inhibit a best adipocyte differentiation. The 3. 10-4,10-5 g / mL concentration of leaf extract can upregulate the expression of Cbfa 1mRNA prompted leaf extract induce MSCs osteogenic differentiation of possible mechanisms.

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