Effects Of Chitosan Membrane Culture On Expressions Of Stemness Related Genes In Mesenchymal Stem Cells

Objective:To investigate the effects of chitosan membrane on mesenchymal stem cells(MSCs)sternness by comparing the related genes expression levels in two different plate cultured MSCs.Method:MSCs were isolated from umbilical cords and cultured on chitosan membrane and in conventional culture condition.Expressions of mRNA of sternness related genes including Nanog,Sox2,c-Myc and Oct-4 were assayed by real-time reverse transcriptase polymerase chain reaction.And the expressions of proteins of sternness related genes were detected by Western Blot method.Result:MSCs cultured on chitosan membrane can form spheroid.The mRNA expression levels of Nanog,Sox2,c-Myc and Oct-4 were 3.77±0.25,2.32±0.18,2.49±0.21 and 2.96±0.23 respectively in chitosan membrane-cultured MSCs.The expression levels were significantly higher than that of conventional culture group.Compared with conventional culture group(Nanog:1.11±0.12;Sox2:0.38±0.06;c-Myc:0.63±0.07;Oct-4:1.26±0.14),protein expressing levels of the four genes increased significantly(Nanog:2.87±0.22;Sox2:0.89±0.08;c-Myc:1.24±0.13;Oct-4:2.71±0.26.all P<0.01).Conclusion:Chitosan membrane-cultured MSCs can form spheroid.The expression levels of sternness related genes in these MSCs were upregulated significantly.MSCs cultured on chitosan membrane can maintain its sternness.