E-GEOD-8023 - Transcription profiling by array of human CD34-positive umbilical cord blood cells after AML1-ETO fusion gene transduction

Status
Released on 15 June 2008, last updated on 30 April 2015
Organism
Homo sapiens
Samples (12)
Array (1)
Protocols (4)
Description
AML1-ETO expression in normal human umbilical cord blood CD34+ cells leads to long-term proliferation of an early self-renewing primitive progenitor cell with multilineage potential and stem cell ability, but these cells do not induce leukemia in immunodeficient mice. This comparative microarray study was initiated to determine the differences in the transcriptome of AML-ETO-expressing CD34+ cells after extended culture in vitro, using normal cord blood cells expanded for 6-8 weeks in vitro and subsequently purified for the CD34+ population as the control comparison. Experiment Overall Design: We have established a culture system whereby we retrovirally transduce human CD34+ cells, obtained from cord blood, with the leukemia fusion gene AML1-ETO. Cells expressing this fusion protein are able to proliferate long-term in vitro in a cytokine dependent manner. AML1-ETO-expressing cord blood cells have a large population of primitive self-renewing CD34+ cells with continued abnormal differentiation. We grow these cells in serum-free conditions using the BIT supplement from Stem Cell Technologies. For the current experiments we used cell cultures that had been proliferating in vitro for 8-12 weeks, in a cytokine cocktail of SCF, TPO, FLT3L, IL-6 all at 20 ng/mL and IL-3 at 10 ng/mL. Control cord blood samples that were CD34 purified were expanded for 5-8 weeks in the same culture media as used for AML1-ETO cells. All samples were magnetically selected for the CD34+ population, returned to culture, and one week later again selected for CD34+ cells and then lysed for RNA isolation.
Experiment types
transcription profiling by array, genetic modification design
Contact
Citation
p53 signaling in response to increased DNA damage sensitizes AML1-ETO cells to stress-induced death. Ondrej Krejci, Mark Wunderlich, Hartmut Geiger, Fu-Sheng Chou, David Schleimer, Michael Jansen, Paul R Andreassen, James C Mulloy.
MIAME
PlatformsProtocolsVariablesProcessedRaw
Files
Investigation descriptionE-GEOD-8023.idf.txt
Sample and data relationshipE-GEOD-8023.sdrf.txt
Raw data (1)E-GEOD-8023.raw.1.zip
Processed data (1)E-GEOD-8023.processed.1.zip
Array designA-AFFY-44.adf.txt
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