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Transcriptome analysis of immediate early events in mRNA-mediated cellular reprogramming of human foreskin fibroblasts

ABSTRACT: In this study human neonatal foreskin fibroblasts (HFF1 and BJ) were lipofected with an equal mixture of human reprogramming factor-encoding mRNAs. The cells were lysed and total RNA was harvested 24 hours post-transfection. Gene regulation was evaluated with respect to corresponding mock-transfected, Lipofectamine-treated control fibroblasts. For comparison, wildtype (un-treated) HFF1 and BJ fibroblasts as well as undifferentiated HFF1- and BJ-derived induced pluripotent stem cells and human embryonic stem cells (generated and maintained in our laboratory (refer to GSE26575) were included in the analysis. Total RNA obtained from untreated human neonatal foreskin fibroblasts, reprogramming factor- and mock(Lipofectamine-treated)-transfected human neonatal fibroblasts, undifferentiated hESCs and iPSCs (derived from human neonatal foreskin fibroblasts).

ORGANISM(S): Homo sapiens

SUBMITTER: Katharina Wolfrum 

PROVIDER: E-GEOD-34943 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

The cytotoxic and immunogenic hurdles associated with non-viral mRNA-mediated reprogramming of human fibroblasts.

Drews Katharina K   Tavernier Geertrui G   Demeester Joseph J   Lehrach Hans H   De Smedt Stefaan C SC   Rejman Joanna J   Adjaye James J  

Biomaterials 20120228 16

Delivery of reprogramming factor-encoding mRNAs by means of lipofection in somatic cells is a desirable method for deriving integration-free iPSCs. However, the lack of reproducibility implies there are major hurdles to overcome before this protocol becomes universally accepted. This study demonstrates the functionality of our in-house synthesized mRNAs expressing the reprogramming factors (OCT4, SOX2, KLF4, c-MYC) within the nucleus of human fibroblasts. However, upon repeated transfections, th  ...[more]

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