Next Generation Sequencing based mRNA sequencing of osteogenic differentiation of human mesenchymal stem cells from hair follicle outer root sheath (MSCORS)

Next Generation Sequencing (NGS)-based mRNA sequencing (RNA-seq) has provided high-throughput measurements OF MRNA COPIES? 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).The cells were subjected to osteogenic differentiation for 1, 2 and 3 weeks. mRNA profiles of differentiated MSCORS afer 1, 2, and 3 weeks 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 differentiated human mesenchymal stem cells from hair follicle outer root sheath after 1 week, 2 weeks and 3 weeks osteogenic differentiation