Combined characterization of microRNA and mRNA profiles delineates early differentiation pathways of CD133+ and CD34+ hematopoietic stem and progenitor cells.
Bottom Line: Using quantitative whole genome miRNA microarray and sequencing-based profiling, we found that between 109 (CD133(+) ) and 216 (CD34(-) CD133(-) ) miRNAs were expressed.In conclusion, the miRNAs expressed differentially between the CD133(+) and CD34(+) CD133(-) cells are involved in inhibition of differentiation, prevention of apoptosis, and cytoskeletal remodeling.These results are highly relevant for stem cell-based therapies with CD133(+) cells and delineate for the first time how the stem cell character of CD133(+) cells is defined by the expression of specific miRNAs.
Affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.Show MeSH
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Mentions: CD133+ cells were magnetically isolated from bone marrow by MACS technology. Subsequently, CD34+CD133− cells were isolated using the negative fraction of the first separation (Fig. 1A). Besides the CD34+CD133− cell population, CD34−CD133− cells (the negative fraction of the second separation) were also analyzed and compared with CD133+ cells to get a general impression of miRNA expression in bone marrow cells. As the main cell type present in the CD34−CD133− fraction are erythrocytes, an additional analysis of CD34−CD133− cells after red blood cell (RBC) lysis was performed (Neg −R). Here, the main cell population is neutrophilic granulocytes .
Affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.