Human Adipose Mesenchymal Stem Cells (hAMSCs) derived Exosomal-miRNA sequencing. Homo sapiens

Exosomes is a type of micro-vesicles having 30-200 nm diameters, secreted by all cells, could be up-taken by neighboring cells, containing many important components including RNAs, proteins, DNA and lipids, serving as efficient vehicles for cancer-stromal communication. NGS was performed to analyse the enrichment of miRNAs in hAMSCs-CM derived exosomes.Exosomal-RNA was isolated from hAMSCs-CM derived exosomes, small RNA libraries were prepared through a series of processes such as adapter ligation, reverse transcription, PCR amplification, and pooled gel purification.Sequence quality was checked by FastQC experiment, unique sequences found for known miRNAs after clustering were verified with miRBase database (www.mirbase.org/) by blast, trimmed read might have considered as miRNA on condition of having 100% identical and whole length sequence of miRNA compare to the ones on miRBase database. Unmatched trimmed read excluded from miRBase were blast to ncRNA database Rfam (http://rfam.sanger.ac.uk/). Unmatched trimmed reads to any of non-miRNA on Rfam database were considered as a potential novel miRNA.Performing NGS of exosomal-RNAs, we identified 141 known miRNA (common in all three samples) and majority of them are involved in different molecular and biological processes including which are regulation of oncogenesis.In conclusion, hAMSCs-CM derived exosomes contains many miRNAs which are potentialy involved in the regulation of oncogenes.