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Proc Natl Acad Sci U S A. 2010 Aug 31;107(35):15443-8. doi: 10.1073/pnas.1009320107. Epub 2010 Aug 11.

Comprehensive microRNA expression profiling of the hematopoietic hierarchy.

Author information

1
Center for High-Throughput Biology, Department of Physics and Astronomy, and Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada V6T 1Z4.

Abstract

The hematopoietic system produces a large number of highly specialized cell types that are derived through a hierarchical differentiation process from a common stem cell population. miRNAs are critical players in orchestrating this differentiation. Here, we report the development and application of a high-throughput microfluidic real-time quantitative PCR (RT-qPCR) approach for generating global miRNA profiles for 27 phenotypically distinct cell populations isolated from normal adult mouse hematopoietic tissues. A total of 80,000 RT-qPCR assays were used to map the landscape of miRNA expression across the hematopoietic hierarchy, including rare progenitor and stem cell populations. We show that miRNA profiles allow for the direct inference of cell lineage relations and functional similarity. Our analysis reveals a close relatedness of the miRNA expression patterns in multipotent progenitors and stem cells, followed by a major reprogramming upon restriction of differentiation potential to a single lineage. The analysis of miRNA expression in single hematopoietic cells further demonstrates that miRNA expression is very tightly regulated within highly purified populations, underscoring the potential of single-cell miRNA profiling for assessing compartment heterogeneity.

PMID:
20702766
PMCID:
PMC2932570
DOI:
10.1073/pnas.1009320107
[Indexed for MEDLINE]
Free PMC Article
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