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Global gene expression analysis of human embryonic stem cells, adult fibroblasts , and CD34+ cord blood (CB) cells before, during, and afer their episomal induction of pluripotency

ABSTRACT: Global gene expression analysis of (a) human embryonic stem cells, (b) adult fibroblasts with and without nucleofection of SOKM, and ( c ) CD34+ cord blood cells at various time points during induction of pluripotency with SOKM, with or without co-culture with bone marrow stromal cells (BMSC).

ORGANISM(S): Homo sapiens

PROVIDER: GSE35027 | GEO | 2012/08/23

SECONDARY ACCESSION(S): PRJNA156183

REPOSITORIES: GEO

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Global gene expression analysis of human embryonic stem cells, adult fibroblasts , and CD34+ cord blood (CB) cells before, during, and afer their episomal induction of pluripotency

Project description:Global gene expression analysis of (a) human embryonic stem cells, (b) adult fibroblasts with and without nucleofection of SOKM, and ( c ) CD34+ cord blood cells at various time points during induction of pluripotency with SOKM, with or without co-culture with bone marrow stromal cells (BMSC). Total RNA was harvested from adult fibroblasts, adult fibroblasts three days after nucleofection with 4F (SOKM), Day -3 unstimulated cord blood (CB) cells, Day 0 growth factor stimulated CB cells, Day +3 CB cells with or without nucleofection at Day zero and with or without Day zero to Day +3 co-culture with bone marrow stromal cells (BMSC), Day 23 CB cells nucleofected on day zero and co-cultured on BMSC from Day Zero to Day +3, and undifferentiated H9 human embryonic stem cells. All experimental conditions repeated in three independent trials, and each replicate at each condition analyzed on an individual microarray.

2012-08-23 | E-GEOD-35027 | biostudies-arrayexpress

Global gene expression analysis of human embryonic stem cells, adult fibroblasts, and CD34+ cord blood (CB) cells before, during, and after generation of stromal primed and non-stromal primed episomal induction of pluripotency

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Project description:In order to investigate the role of CD34 antigen in haematopoietic commitment, we silenced the CD34 gene expression in CD34+ stem/progenitor cells using a siRNA approach. Experiment Overall Design: To maximize siRNA transfection efficiency, we utilized the NucleofectorTM technology (Amaxa). CD34+ cells were transfected with a mixture of 4 siRNAs targeting CD34 mRNA and with a non-targeting siRNA as a negative control. The expression level of CD34 antigen on control cells (MOCK and negative control treated cells) and CD34siRNA treated cells was assessed by immunofluorescence analysis at 24 and 48h post-nucleofection.

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