Next Generation Sequencing-based on mRNA sequencing of human adipose mesenchymal stem cells submitted to osteogenic differentiation

Next Generation Sequencing (NGS)-based mRNA sequencing (RNA-seq) has provided high-throughput measurements to analyze the transcriptome of differentiating adipose mesenchymal stem 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. ADMSC were isolated from human adipose tissue, cultured and characterized as mesenchymal stem cells (MSC) according to the minimum criteria for defining multipotent MSC from the International Society for Cellular Therapy (ISCT). After osteogenic differentiation of the ADMSC, mRNA profiles of ADMSC differentiated for 1 week, 2 weeks and 3 weeks were quality controlled, and generated by deep sequencing 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 differentiated human adipose-derived mesenchymal stem cells after 1 week, 2 weeks and 3 weeks osteogenic differentiation