mRNA expression profiles 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 gene expression comparison between the substate 1 and substate 2 cells, DNA 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. The total mRNAs were extracted from substate 1 and substate 2 cells by using ISOGEN (NIPPON GENE). DNA microarray were performed by a SurePrint G3 Human GE Microarray Kit, 8 x 60 K Ver. 3 (Agilent, G4851C).