Basic & Clinical Medicine ›› 2013, Vol. 33 ›› Issue (10): 1229-1234.

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In Vitro synthesized modified mRNA can enter the hUC-MSCs

  

  • Received:2012-11-19 Revised:2013-01-24 Online:2013-10-05 Published:2013-09-25

Abstract: Objective To investigate the stability and efficiency of the in vitro synthesized modified mRNA, when it's transfected into the human umbilical cord mesenchymal stem cells(hUC-MSCs), so to establish a platform of using mRNA to induce the hUC-MSC differentiate into other cells for cell therapy. Methods To get the plasmid construct as the template for mRNA synthesis and synthesize the mRNA of eGFP. When the modified mRNA was transfected into the hUC-MSCs, using flow cytometry to analyze the transfect efficiency and determine the best transfect doses also to find the half-life time of mRNA. The same method was used to synthesize the mRNA of Pdx1 and to transfect it into the hUC-MSCs, then to measure the transfect efficiency by immunoflurescence. Result The in vitro synthesized modified mRNA of eGFP can enter the hUC-MSCs efficiently, the best doses for transfection is 1.5μg/mL and the mRNA can stay in the cell stably for 3 days. The in vitro modified mRNA of Pdx1 also can enter the hUC-MSCs. Conclusion In vitro synthesized modified mRNA has good stability and can enter the cells and translate into protein.

Key words: Modified mRNA, hUC-MSC, Transfection efficiency