Abstract
Aim:Nonviral induced pluripotent stem cell (IPSC) reprogramming is not efficient without the oncogenes, Myc and Lin28. We describe a robust Myc and Lin28-free IPSC reprogramming approach using reprogramming molecules.Methods:IPSC colony formation was compared in the presence and absence of Myc and Lin28 by the mixture of reprogramming molecules and episomal vectors.Results:While more colonies were observed in cultures transfected with the aforementioned oncogenes, the Myc and Lin28-free method achieved the same reprogramming efficiency as reports that used these oncogenes. Further, all colonies were fully reprogrammed based on expression of SSEA4, even in the absence of Myc and Lin28.Conclusion:This approach satisfies an important regulatory pathway for developing IPSC cell therapies with lower clinical risk.
Highlights
We describe a robust Myc and Lin28-free induced pluripotent stem cell (IPSC) reprogramming approach using reprogramming molecules
We report an IPSC reprogramming method, which uses a combinatorial approach of reprogramming molecule enhancers and a mixture of episomal vectors that are free of Myc and Lin28
We observed at most one colony formation for 100,000 input cells in the absence of reprogramming molecules regardless of whether cells were transfected in the presence or absence of Myc and Lin28 transcriptional factors, which is in agreement with other reports [16]
Summary
Nonviral induced pluripotent stem cell (IPSC) reprogramming is not efficient without the oncogenes, Myc and Lin
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