Differentiation Of Embryonic Stem Cells Into Vascular Smooth Muscle Cells Promoted By MiRNA-145

Objective: In this study,mi RNA-145 was used as inducer in combination with other inducers to induce ESCs to differentiate into VSMCs,and a scheme of mi RNA-145 to promote ESCs to differentiate into VSMCs was initially established.Finally,in the optimal induction scheme,KLF4 was overexpressed,and the mechanism of mi RNA-145 promoting ESCs differentiation into VSMCs was preliminarily studied.This will lay a foundation for further research on the regulation mechanism and the establishment of a faster and more efficient ESC-VSMCs induction scheme.Methods: 1.During ESCs induced differentiation into VSMCs,the expression of mi RNA-145,KLF4 and VSMCs specific marker proteins were detected.2.mi RNA-145 was used as inducer and combined with different inducers to establish the best induced differentiation scheme.On the basis of this scheme,KLF4 was overexpressed to observe the expression of VSMCs specific marker protein and the contraction function of induced differentiated cells.Results:1.Experimental Study on Expression of mi RNA-145 in Inducing Differentiation of Embryonic Stem Cells into Vascular Smooth Muscle Cells.(1)ESC-VSMCs was successfully induced.With the induction,the clonal-like cells,which showed close contact under light microscope,gradually changed to long spindle shape.(2)In different induction stages,Q-PCR showed that mi RNA-145 and VSMCs specific markers ?-SMA,Calponin,SM22? changed dynamically,and KLF4 expression decreased on the days 1-3.The expression of mi RNA-145 increased significantly from day 3 to day 6and reached a peak.The specific marker genes showed the same change trend.(3)The specific marker proteins ?-SMA,Calponin and SM22? in ESC-VSMCs were all expressed positively by immunofluorescence staining.2.Experimental Study on Expression of mi RNA-145 in Inducing Differentiation of Embryonic Stem Cells into Vascular Smooth Muscle Cells.(1)Different combinations of mi RNA-145,Activin A and PDGF-BB can be used as inducers to induce VSMCs that both express specific marker proteins and have contractile function,but different groups have differences,of which mi RNA-145 combined with PDGF-BB has the best induction efficiency.(2)Mi RNA-145 may promote stem cells to differentiate into VSMCs by inhibiting KLF4,especially in the process of promoting intermediate lineage cells to differentiate into VSMCs.Conclusion: 1.Different combinations of mi RNA-145,Activin A and PDGF-BB can be used as inducers to induce VSMCs that not only express specific marker proteins but also have contractile function.However,different groups have differences,and mi RNA-145 combined with PDGF-BB has the best induction efficiency.2.mi RNA-145 may promote the induced differentiation of stem cells into VSMCs by inhibiting KLF4,especially in the process of promoting the differentiation of intermediate lineage cells into VSMCs.