| Objective:In this study, adult rat bone marrow cells by density gradient centrifugation combined with adherent method isolated BMSCs were cultured and passaged,and Then injected rat model of denervated muscle nerve ends, were observed bone marrow mesenchymal stem cells on prevention and treatment of denervated skeletal muscle atrophy and provide a new theoretical basis for clinical treatment.Methods:1 SD rats 1, femur and tibia bone marrow cells under sterile conditions, the bone marrow cells isolated by percoll solution (specific gravity 1.077) isolated bone marrow mesenchymal stem cells in mononuclear cells, and then inoculated with a single cell suspension DMEM 15% calf serum medium was cultured, purified and amplified. Draw out the MSCs growth curve was detected by immunohistochemistry stem cells CD44, CD45 surface markers.2 The rat bone marrow mesenchymal stem cells was cultured and separated using stick wall filtering method.36 healthy SD rats were cut off the left sciatic nerve, caused the nerve defect, were randomly divided into Observation group and control group.The Observation group on the left side of the gastrocnemius muscle injection of 1.5 × 105 cells/ml bone marrow mesenchymal stem cells 0.2ml, the control group on the left side of the gastrocnemius muscle intramuscular injection of the same dose of sugar DMEM Broth, Respectively, in2、4、6 weeks after the general observation on the two groups of rats were detected muscle wet weight, muscle fiber diameter and cross-sectional area, nerve index (SFI), Immunohistochemistry was used to detect muscle tissue Bcl-2, Bax expression change.Results:Using the density gradient centrifugation and adherent culture could be effectively separated and purified BMSCs, cells were cultured stem cells with typical morphology, After the Passaging of cell proliferation faster. BMSCs were positive for CD44 expression markers, hematopoietic stem cell marker CD45 negative expression.2、4、 6 weeks After establishing the model, The experimental group rats, the muscle wet weight, the muscle fiber diameter and cross-sectional area were significantly higher than the control group, SFI was significantly higher than that of control group, the difference was statistically significant (P<0.05).2,4,6 weeks postoperatively, in the observation group of muscle tissue Bcl-2 positive cells expressing intensity were higher than the control group, Bax positive cells expressing intensity are lower than the control group, the differences were statistically significant (P<0.05).Conclusions:1.By density gradient centrifugation and adherent culture method can effectively separate the BMSCs of high purity, and has a good growth rate, in vitro after in vivo transplantation of BMSCs can be carried out, and play a role in local damage microenvironment.2.BMSCs were injected into the loss of skeletal muscle nerve newborn can promote nerve cell maturation, improve transplant nerve function, delaying denervation of skeletal muscle atrophy. |
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