Basic & Clinical Medicine ›› 2012, Vol. 32 ›› Issue (9): 1030-1035.

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Induction of human fibroblast cells to pluripotent stem cells and differentiation into cardiomyocytes

  

  • Received:2011-06-13 Revised:2011-12-29 Online:2012-09-05 Published:2012-08-28
  • Supported by:
    Singapore National Research Foundation;Grant of Department of Education, Henan Province

Abstract: Abstract: Objective To increase the transduction efficiency of iPSC formation, decrease the time schedule and avoid the possibility of exogene introduction. Methods Human fibroblast cells were infected with retrovirus of Oct4, Sox2, Klf4 and c-myc which were packaged by GP2-293 cells. During the transduction, small molecules of SB431542,PD0325901 and thiazovivin were added into hESC medium, until iPSC colonies generated big enough to be cut and paste. The stable iPSC colonies were detected with the expression of ESC protein markers, cardiomyocytes differentiation and karotyping analysis. Results On day 13 small iPSC colonies were shown up, and on day 20 they were ready to be cut and transferred to cell culture dishes. The reprogramming period was shortened for 10 days, and the efficiency was increased by 12 times. The iPSC shared protein expression with hESC on Oct4,SSEA3,Tra-1-60 and Tra-1-81. As well, both iPSC and hESC were differentiated into cardiomyocytes successfully with similar MEA data. And both showed normal karotyping results. Conclusion The iPSC reprogramming system established are efficient and time-saving for study on disease-specific model and holding clinic poteintial.

Key words: Key words: human induced pluripotent stem cells, transduction efficiency, time schedule, cardiomyocytes differentiation

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