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Mary Ann Liebert, Stem Cells and Development, 19(22), p. 2641-2654

DOI: 10.1089/scd.2013.0040

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Defining the Diversity of Phenotypic Respecification Using Multiple Cell Lines and Reprogramming Regimens

This paper is available in a repository.
This paper is available in a repository.

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Abstract

To better understand the basis of variation in cellular reprogramming, we performed experiments with two primary objectives: first, to determine the degree of difference, if any, in reprogramming efficiency among cells lines of similar type after accounting for technical variables, and second, to compare the efficiency of conversion of multiple similar cell lines to two separate reprogramming regimens - induced neurons and induced skeletal muscle. Using two reprogramming regimens it could be determined if converted cells likely are derived from a distinct subpopulation generally susceptible to reprogramming or are derived from cells with independent capacity for respecification to a given phenotype. Our results indicated that when technical components of the reprogramming regimen were accounted for, reprogramming efficiency was reproducible within a given primary fibroblast line but varied dramatically between lines. The disparity in reprogramming efficiency between lines was of sufficient magnitude to account for some discrepancies in published results. We also found that the efficiency of conversion to one phenotype was not predictive of reprogramming to the alternate phenotype, suggesting that capacity for reprogramming does not arise from a specific subpopulation with a generally "weak grip" on cellular identity. Our findings suggest that parallel testing of multiple cell lines from several sources may be needed to accurately assess the efficiency of direct reprogramming procedures, and that testing a larger number of fibroblast lines -- even lines with similar origins -- is likely the most direct means of improving reprogramming efficiency.