| Backround and Objective:Bone marrow mesenchymal stem cells (MSCs) are bone marrow nonhematopoi- etic stem cells with self-replication ability and multilineage differentiation potential. Under different conditions can be induced to differentiate into multiple cell lines, including osteoblasts, fibroblasts, cartilage cells, fat cells, muscle cells, endothelial cells and neural cells.Only the traditional agent-induced osteogenesis alone or the role of adipogenic differentiation, which has a limited application. Extracting solution from Eucommia ulmoides oliv as an induction agent, has to increase bone density and inhibit bone resorption and regulate the efficacy of the role of bone metabolism, could promote osteoblast proliferation and alkaline phosphatase secretion, while inhibiting fat cell differentiation to the role of .Experimental Investigation of the Eucommia leaf extract induced sheep MSCs to osteoblast differentiation at the same time, confirmed its inhibitory differentiation of MSCs into the role of fat.Contents and methods of research:1. The use of the whole bone marrow in vitro isolation and culture of goat MSCs, conventional methods of mass culture, flow cytometry CD29, CD44 and CD105 to identify MSCs.2. Extracting solution from Eucommia ulmoides oliv on the proliferation of MSCs and osteogenic differentiation. Detection of ALP, calcium staining nodules in order to observe the effect of the promotion of osteogenic differentiation.3. Extracting solution from Eucommia ulmoides oliv MSCs inhibit the differentiation into fat. Line oil red-O staining to observe the effect of inhibiting differentiation into fat.4. The level of detection from the cell extracting solution from Eucommia ulmoides oliv osteogenic differentiation of MSCs after expression of Cbfa lmRNA using automatic gel image analysis analyzer to observe its effects to promote osteoblast differentiation. Results:1. Bone marrow culture-wide (that is, law adherent) Isolation and Purification of goat MSCs by trypsin digestion for liquid and gradually purified after passage, method is simple and practical.2. With control group, the osteoblast cells induced by an increase in alkaline phospha tase synthesis, the formation of mineralized nodules, and the traditional agent-induced osteogenesis + extracting solution from Eucommia ulmoides oliv group ,10-4 ,10-5g / mL extracting solution from Eucommia ulmoides oliv osteoinduction case group supe rior to the traditional agent-induced osteogenesis Group ,10-6g / mL extracting solut ion from Eucommia ulmoides oliv group.3. Oil red O staining showed that adding extracts of extracting solution from Eucommia ulmoides oliv of the adipogenic induction group, the number of lipid droplets significantly less than traditional adipogenic induction agent group and blank control group to 10-4,10-5g /mL extracting solution from Eucommia ulmoides oliv group best.4. RT-PCR showed that the osteoblasts induced by high-performance group were amplified the 171bp fragment of gene transcription Cbfa 1mRNA and 10-4,10-5g / mL concentration of extracting solution from Eucommia ulmoides oliv in osteoblasts the expression level of Cbfa 1mRNA the difference was not significant (P> 0.05).Conclusion:1. Extracting solution from Eucommia ulmoides oliv can be induced by in vitro culture of sheep separation and purification of bone marrow mesenchymal stem cells to the direction of osteoblast differentiation and proliferation, while inhibiting their differentiation to adipocytes, the role of two-way adjustment.2. Extracting solution from Eucommia ulmoides oliv to 10-4,10-5g / mL concentration of osteoblasts and inhibit differentiation into fat best.3. 10-4,10-5 g / mL concentration of extracting solution from Eucommia ulmoides oliv can increase the expression of Cbfa1mRNA, suggesting extracting solution from Eucommia ulmoides oliv-induced osteogenic differentiation of MSCs may be a mechanism. |
PDF Full Text Request