Next Generation Sequencing based mRNA sequencing of human mesenchymal stem cells from hair follicle outer root sheath

Next Generation Sequencing (NGS) based mRNA sequencing (RNA-seq) has provided high-throughput measurements to analyze the transcriptome of cells. We report the comprehensive transcriptome profiling of primarily isolated human mesenchymal stem cells from hair follicle outer root sheath (MSCORS) in series cultivation passages using NGS-based RNA-seq method. Human anagen hair follicles were non-invasively plucked from scalp of healthy donors, cultivated on a Transwell membrane, from which the MSCORS were isolated and subcultured onto normal culture flask. Attached and proliferating MSCORS were 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 MSCORS in p0, p1 and p2 were quality controlled, and generated by deep sequencing, in triplicate, using Illumina PE150. 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). mRNA profiles of human mesenchymal stem cells from hair follicle outer root sheath (MSCORS) in p0, p1 and p2