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MiRNA expression of the isolated human platelet-derived microparticle

ABSTRACT: To analysis of potential functional components in PMPs to miRNAs, we have employed miRNA microarray expression profiling as a discovery platform to identified the miRNA content of PMPs.Human cord blood from healthy donors was used to isolated platelets and platelet-derived microparticles (PMPs) were isolated from activated platelets. First, for platelet isolation and purification, human platelets were isolated from the supernatant of umbilical cord blood after Ficoll-Hypaque density gradient centrifugation and pelleted by centrifugation at 2,000 × g for 20 min at RT. Second, the platelets were resuspended in saline (pH=7.4) containing 1 mM calcium and 1 U per mL thrombin for 15 min at 37°C. The PMPs were released into the supernatant. Contaminating remnant platelets were removed by centrifugation at 2,000 × g for 20 min at 4°C and filtered through a 0.8 μm filter unit (Millipore, America). Then, the supernatants containing PMPs were centrifuged at 20,000 × g for 120 min at 4°C.182 miRNAs were detected in the isolated human PMPs from five independent donors, although individual differences in miRNA expression were also observed. Thereafter, these miRNAs were ranked by expression level, and those expressed above the median level from each unique subject were considered to be abundantly expressed miRNAs. The intersection of abundantly expressed miRNAs in PMPs from each donor yielded 48 commonly expressed miRNAs in PMPs. Among these 48 miRNAs, miR-4454 ranked number 1, and its level was consistent in all samples examined.

ORGANISM(S): Homo sapiens

PROVIDER: GSE152078 | GEO | 2020/09/07

REPOSITORIES: GEO

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