|
Studies have shown that muscle-derived stem cells (muscle derived stem cells, MDSCs) may be a novel skeletal muscle-derived adult stem cells, tissue engineering and gene therapy has broad application prospects. Currently, MDSCs research there are many hot issues, and how to get to meet experiments and future clinical required MDSCs is an urgent need to solve the problem. Many scholars believe that the use of growth factor amplification method is a good choice, The Study is underway. Of the study carried out in this paper aims to further explore of MDSCs the amplification conditions, experimental basis for the research and application. The isolated rat MDSCs in this experiment by enzyme digestion by density gradient centrifugation and differential adherence purified, and then will get MDSCs cultured in vitro, and by the immunocytochemistry Act (desmin, Sca-1) obtained The cells were identified. Finally, the MTT assay at different concentrations (6.25,12.5,25,50,100 ng / ml), IGF-1 and CNTF alone 24h, 48h, 72h and 96h light absorption value of 96h, and the two combined effects (100ng/ml) (OD). The resulting data were analyzed through SPSS13 statistical package, data are expressed as mean ± standard deviation, the groups were compared using analysis of variance, pairwise comparisons using Tukey's method, the P value lt; 0.05 as statistically significant. The results show that: the experimental group (were added to different concentrations of IGF-1 and CNTF) absorbance value (OD) were higher than the negative control group, but in 6.25ng/ml group, CNTF and IGF-1 on MDSCs promote proliferative effect was not significant. In 12.5ng/ml group, two factors on proliferation effect of MDSCs significantly increased (P lt; 0.05);, but no significant difference in both pro-proliferative effect. 25ng/ml group, CNTF and IGF-1 on MDSCs promote the proliferation not only further enhanced between the existence of a significant difference (P lt; 0.05). 50ng and 100ng/ml group, CNTF and IGF-1 promote MDSCs proliferative effects were close to their peak and plateau phenomenon. With the prolonged incubation time, the light absorption values ??of the different points in time is significant difference. The negative control group as reference, significantly with sex and promote proliferation effect (P lt; 0.05) the in vitro culture, respectively, as: 96h (IGF-1 group), 72h (of CNTF group), 48h (IGF-1 of CNTF group ). The 24h experimental factors promoting value-added effect is not obvious, there is no significant difference. During the 48h-96h, the combined group (IGF-1 and CNTF) promote of MDSCs the proliferation effect most significant difference (P lt; 0.05), and other experimental groups exist immunocytochemistry results show: culture 4d was Sca-1 positive reaction the MDSCs more, the positive rate of 90%. Desmin positive MDSCs smaller number. This experiment suggests: CNTF and IGF-1 have rats promoted the MDSCs proliferation effect, and there is the phenomenon of synergies. But alone, both promote the effectual time of MDSCs proliferation significant differences, and 6.25ng/ml 12.5ng/ml 25ng/ml, 50ng/ml group, promoting MDSCs proliferation in a concentration-dependent manner. Relevant mechanism of action is still unclear.
|