Effect Of The Conditioned Medium Of HAMSCs On Human Fibroblasts

Objective: To isolate and culture human amnion mesenchymal stem cells(hAMSCs), and explore the effect of the conditioned medium of hAMSCs(hAMSC-CM) on human fibroblasts. Methods: We isolated from human amnion tissue and culture hAMSCs, and use vimentin immunofluorescence and flow cytometry to identify hAMSCs. Then the conditioned medium of hAMSCs was used to culture human fibroblasts and measured by ELISA kit. The cells counting by the CCK8 method was to determine the proliferation of human fibroblasts. Annexin V/PI was used to investigate the apoptosis rate of human fibroblasts; the cell migration was measured by in intro wound-healing assay.Then we use statistical software SPSS18.0 experimental data one-way ANOVA analysis and LSD pairwise comparison, α=0.05. Results: hAMSCs was derived by culture. The third generation of hAMSCs expression of vimentin was strongly positive. The positive rates of CD105, CD73 and CD45 CD90 in the third generation hAMSCs cell surface markers were:(77. 1 ± 2.3)%,(65.8 ±1.5)%,(61.7 ± 1.5)%,(6. 0 ± 1.2)%. The contents of IGF-1, VEGF, EGF, bFGF in hAMSC-CM were: 11.69±1.04 pg/mL, 316.47±67.94 pg/mL, 6.12±0.41pg/mL, 1.49±0.05 pg/mL. Compared with those in control group, the proliferation and migration in hAMSC-CM group were significantly higher than that in control group(P <0.05); and the apoptosis rate of fibroblasts in hAMSC-CM group was also found to lower than that control group(P <0.05).Conclusion: The h AMSC-CM could apparently promote human fibroblasts on proliferation, migration and inhibit apoptosis.