RNA sequencing to compare gene expession in hepatic stellate cells on a soft (400 Pa) and stiff (25000 Pa) polyacrylamide hydrogels. Homo sapiens

Primary human hepatic stellate cells (HSCs) isolated from healthy patients were purchased from Sciencell. They were seeded on either 400 Pa or 25000 Pa polyacrylamide hydrogels for 2 days and collected for RNA isolation. The goal of this study is to determine genes transcriptionally regulated by a stiff matrix. 2 RNA samples of HSCs on 400 Pa and 3 RNA samples of HSCs on 25600 Pa were sent to the University of Minnesota Genomics Center for RNA sequencing. Overall design: 2 RNA samples isolated from HSC on 400 Pa stiffness and 3 from HSCs on 25600 Pa stiffness. They were converted to Illumina sequencing libraries using Illumina’s Truseq Stranded mRNA Sample Preparation Kit. Truseq libraries were then subjected to cluster using Illumina cBot instrument and sequencing using HiSeq2500