Microarray Experiments

Cells samples including hUC-MSCs and hUC-MSCs after hepatic differentiation for 2 days, 6 days, 10 days, 14 days, 22 days and 26 days were collected for microRNA expression analysis with Agilent human miRNA (8*60K) V16.0. Microarray experiments including RNA extraction and purification, RNA labeling, array hybridization and data acquisition were performed at Shanghai Biochip Company according to the protocols for the Agilent miRNA microarray system. Total RNA was extracted and purified with a mirVana™ miRNA Isolation Kit (Ambion, Austin, TX, US) and checked for a RIN number to inspect RNA integration with an Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, US). miRNA molecules in the total RNA were labeled with a miRNA Complete Labeling and Hyb Kit (Agilent technologies). Each slide was hybridized with 100 ng Cy3-labeled RNA with a miRNA Complete Labeling and Hyb Kit (Agilent Technologies) in a hybridization oven (Agilent Technologies) at 55°C, and 20 rpm for 20 hours. After hybridization, the slides were washed in staining dishes (Thermo Shandon, Waltham, MA, US) with a Gene Expression Wash Buffer Kit (Agilent Technologies).