Analysis of differentially expressed microRNA of TNF-α-stimulated mesenchymal stem cells and exosomes from their culture supernatant

Archives of Medical Science : AMS
Hualin MaXinzhou Zhang

Abstract

To analyze the microRNA expression of tumor necrosi factor α (TNF-α) stimulated mesenchymal stem cells (MSCs) and exosomes from their culture supernatant. TNF-α (20 ng/ml) was used to stimulate MSCs, which were then regarded as TNF-α cells (TC), while unstimulated cells were the normal control cells (NCC). MSCs and their culture supernatant were harvested after 48 h. Subsequently, exosomes were isolated from culture supernatants with ExoQuick-TC and were divided into two groups, TNF-α exosomes (TE) and normal control exosomes (NCE). Then, the microRNAs were measured by high-throughput sequencing and the results were differentially analyzed. Finally, the correlation of the target genes corresponding to differently expressed microRNAs was analyzed by gene ontology (GO) and KEGG pathway analysis. High-throughput sequencing showed that the cellular compartment (TC vs. NCC) had 280 microRNAs. miR-146a-5p was a uniquely up-regulated microRNA (p < 0.001) and the most significantly down-regulated microRNA among the 279 microRNAs included was miR-150-5p (p < 0.001). There were 180 differentially expressed microRNAs in the exosome compartment (TE vs. NCE), where miR-146-5p (p < 0.001) was one of 176 upregulated microRNAs and miR-203b-5p ...Continue Reading

Citations

Nov 10, 2018·Expert Review of Clinical Immunology·Karl J Lackner, Nadine Müller-Calleja
Apr 13, 2021·Frontiers in Immunology·Yves-Marie PersDanièle Noël
Jul 24, 2021·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Ying-Jie ZhaoJing Li

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Methods Mentioned

BETA
dynamic light scattering
electrophoresis
PCR
electron microscopy
Transmission electron microscopy

Software Mentioned

TargetScan
SOLiD
GenMAPP
RNA2MAP
DEGseq
SOLID system
miRWalk

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