Transcriptomics

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Transcription profiling of human megakaryocytic differentiation primary CD34+ cell culture


ABSTRACT: Little is known about the global transcriptional program underlying megakaryocytic (Mk) differentiation, maturation, and apoptosis. Using DNA microarrays and Q-RT-PCR, we examined the transcriptional profile of Mk differentiation human CD34-positive hematopoietic stem and progenitor cells cultured with thrombopoietin, interleukin-3, and Flt3-ligand. The goal this study was to identify genes involved in the various facets of Mk differentiation including commitment, polyploidization, proplatelet formation, and apoptosis. Experiment Overall Design: G-CSF mobilized peripheral blood CD34-positive cells were cultured with TPO, IL-3, and Flt3-L to induce Mk differentiation. Samples prior to day 5, including uncultured starting cells, were analyzed directly, whereas samples after and including day 5 were positively selected for CD41a expression immediately prior to RNA isolation. Three biological replicate experiments were analyzed and approximately one-half of the samples from each experiment were technically replicated. Hybridizations were performed in a reference design with all samples labeled with Cy3 and a reference RNA pool labeled with Cy5.

ORGANISM(S): Homo sapiens  

SUBMITTER: Eleftherios T. Papoutsakis  

PROVIDER: E-GEOD-3839 | ArrayExpress | 2008-06-12

SECONDARY ACCESSION(S): GSE3839PRJNA104987

REPOSITORIES: GEO, ArrayExpress

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Publications

Comparative, genome-scale transcriptional analysis of CHRF-288-11 and primary human megakaryocytic cell cultures provides novel insights into lineage-specific differentiation.

Fuhrken Peter G PG   Chen Chi C   Miller William M WM   Papoutsakis Eleftherios T ET  

Experimental hematology 20070301 3


OBJECTIVES: Little is known about the transcriptional events underlying megakaryocytic (Mk) differentiation. We sought to identify genes and pathways previously unassociated with megakaryopoiesis and to evaluate the CHRF-288-11 (CHRF) megakaryoblastic cell line as a model system for investigating megakaryopoiesis. METHODS: Using DNA microarrays, Q-RT-PCR, and protein-level assays, we compared the dynamic gene expression pattern of phorbol ester-induced differentiation of CHRF cells to cytokine-i  ...[more]

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