Comprehensive Profiling of miRNAs in Stem Cells Using Microfluidics
MicroRNAs (miRNAs) have been shown to be key regulators of hematopoiesis. However, the detailed profile of miRNA expression throughout hematopoietic development has not been charted, owing primarily to challenges in obtaining highly purified samples of stem and progenitor cells as well as profiling hundreds of miRNAs from low cell numbers. To overcome these limitations, the new technical poster describes how microfluidic technologies were combined with a 288-plex real time PCR approach, allowing the detection of 288 miRNAs in small numbers (c. 3000) of cells.
The poster describes how over twenty unique murine hematopoietic cell populations were isolated through FACS sorting, including hematopoietic stem cells (HSCs), myeloid and lymphoid progenitor cells and mature lineage populations. Several thousand nanolitre volume quantitative PCR reactions were performed using Fluidigm BioMark™ 48.48 Dynamic Arrays and miRNA specific TaqMan® probes. For each miRNA, synthetic miRNA standards were used to determine the absolute number of miRNA molecules per cell. 133 of the 288 miRNAs tested were detected across all cell populations.
The researchers conclude that Fluidigm's dynamic array provides a rapid, sensitive and cost-effective method for miRNA profiling in rare cell populations. The sensitivity of Fluidigm's dynamic array is shown to detect heterogeneity in miRNA expression at the single cell level in purified populations.
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