Next Generation Sequencing-based mRNA sequencing of human adipose mesenchymal stem cells

Next Generation Sequencing (NGS)-based mRNA sequencing (RNA-seq) has provided high-throughput measurements to analyze the transcriptome of cells. We are reporting comprehensive transcriptome profiling of primarily isolated human adipose-derived mesenchymal stem cells (ADMSC) throughout consecutive culture passages using NGS-based RNA-seq method. Human adipose tissue was obtained from operative remains and and digested in collagenase X. ADMSC were isolated, cultivated and characterized as mesenchymal stem cells (MSC) according to the minimum criteria for defining multipotent MSC from the International Society for Cellular Therapy (ISCT). mRNA profiles of ADMSC in p0, p1 and p2 were quality controlled, and generated by deep sequencing, in triplicate, using Illumina PE150 kits. Paired-end raw data were pre-processed using a published protocol. HISAT2 (Hierarchical Indexing for Spliced Alignment of Transcripts) was used to align reference genome. HTSeq was used to calculate Reads Count, and gene abundence was determined by analyzing FPKM (fragments per kilobase of exon model per million reads mapped). Overall design: mRNA profiles of human adipose-derived mesenchymal stem cells in passage 0, passage 1 and passage 2