Effects Of Exogenous BFGF And CNTF On The Proliferation Of MDSCs Of Rat

At present, there are plenty hot spots about the MDSCs research. The purifying, amplifying, and directional differentiation of vitro cell culture is still under exploration. Special attention usually has been paid to the effects of the biological activity factor on the stem cell proliferation while the appropriate cell culture conditions are mentioned. bFGF and CNTF, as a member of biological activity factors, are of great importance both for its biological activity and effectiveness in the basic and clinical researches.In this thesis, researches and experiments have been designed to explore the adult stem cell culture conditions, which may lay the basic experimental foundation for the future study and application. MDSCs of rat has been isolated through the enzymatic digestion, then purified by density gradient centrifugation and differential adhesion and vitro and cultured. The MTT colorimetric has been employed to assay the effects of bFGF with different concentrations (6.25, 12.50, 25.00, 50.00, 100.00ng/ml), CNTF using alone 96h, and the role of the above two combined with 100.0ng/ml, in 24h, 48h, 72h, and 96h on the proliferation of MDSCs. The obtained cells have been identified with the immunocytochemical method (desmin, Sca-1). The OD of the MTT shows:1. Compared with the negative control group, there is a significant proliferation of MDSCs in all the other experimental groups (P<0.05), except the 6.25 (ng/ml). Among the 6.25ng/ml to 50.00ng/ml, MDSCs proliferation significantly enhances with the increased concentrations of bFGF and CNTF (P<0.05).2. There is no significant difference in the MDSCs proliferation between bFGF and CNTF among the 6.25ng/ml to 50.00ng/ml (P>0.05). With its concentration increased to 50.00ng/ml, the bFGF group is significantly lower than CNTF group in the MDSCs proliferation, and the significant differences exists (P<0.05). However, the proliferation effects of the 50.00ng/m group is close to capacity, and there is no significant statistical difference when compared with the 100.00ng/ml group (P>0.05).3. The proliferation effects of the bFGF and CNTF becomes clear with the extension of the culture time, while there is significant difference in the proliferation of effectual time. Compared with the negative control group, the significant proliferative effect of time presents at bFGF alone 96 h, CNTF alone 72h, combined 48h, respectively (P <0.05). At 24h in the experimental groups, even though the number of MDSCs increases, but there is no significant difference (P >0.05).4. At 48h-96h period, the proliferation effects is of the strongest in bFGF and CNTF combined group, and there is significant difference exists when compared with the other experimental groups (P <0.05).Note: Both of the bFGF and CNTF can promote the vitro cultured rat MDSCs proliferation, and there is a synergy in the proliferation. However, bFGF and CNTF to promote the proliferation of MDSCs is with appearent different time, and at 12.5-50ng/ml period, the role of promoting the proliferation of MDSCs also appears a dose-dependent manner.