| Objective Study of mesenchymal stem cells(MSCs)combined with endothelial progenitor cells(EPCs)to promote tissue engineering vascularization in vitro.Methods1.Isolation and culture of four to six weeks murine bone marrow MSCs and EPCs;2.Morphological characteristics of MSCs and EPCs were observed by microscopy;3.MSCs and EPCs surface markers were detected by flow cytometry;4.Handle the non-woven polyglycolic acid(PGA);5.Inoculation cells: The experiment was divided into MSC group,EPCs group,MSCs+EPCs group,Take the third generation of MSCs and EPCs,respectively,by 3 × 106 cells inoculated on a non-woven polyglycolic acid(PGA)on the culture in vitro for 8 weeks(MSCs+EPCs group were inoculated with a ratio of 1: 1 in 1.5 × 106 cells);6.Cell-scaffold complex sections were stained with HE for histomorphological observation.Immunohistochemical staining to evaluate the expression of CD31 in each experimental group;7.Data was processed and statistically analyzed by SPSS 17.0.Results1.MSCs and EPCs can be effectively separated and amplified by whole bone marrow adherent method combined with differential adhesion method.Cell surface markers were detected by flow cytometry,the third generation of MSCs highly expressed Sca-1,CD29 and CD44,which met the characteristics of MSCs,the third generation of EPCs high expression of CD309,CD34 and CD133,which met the characteristics of EPCs.2.Histological observation showed that the number of cells and morphology of MSCs+EPCs group were better than those of MSCs group and EPCs group,and the metabolites of PGA were almost completely degraded.3.Immunohistochemistry showed that the expression of CD31 in MSCs+EPCs group were higher than those in EPCs group,and EPCs group was higher than that in MSCs group,the difference was statistically significant(P<0.0001).Conclusion MSCs combined with EPCs can promote the vascularization of tissue engineering in vitro,and its proliferation and angiogenesis are higher than that in single culture group. |
PDF Full Text Request