Expression profiles of miRNA derived from exosomes for ectoderm-biased substate of human pluripotent stem cells.

We established two different substates in human ES cell line, H9 cells, by changing the extracellular matrices. To examine a global miRNA expression comparison between the substate 1 and substate 2 cells, miRNA microarray was performed. Human ES cell line, H9 cells, was maintained on Vitronectin XF (STEMCELL Technologies) in TeSR-E8 (STEMCELL Technologies) with feeder free conditions. We changed the extracellular matrices from Vitronectin to Matrigel (BD Biosciences). The H9 cells cultured on Matrigel contained two different substates. Exosomes in the cell culture medium were precipitated by ExoQuick-TC (System Biosciences, EXOTC50A-1). The miRNAs of substate 1 and substate 2 were extracted from the exosomes by using miRNeasy Mini Kit (QIAGEN, 217004). miRNA microarray were performed by a SurePrint G3 miRNA Microarray Kit, 8 x 60K (Agilent, G4872A).