Genome-wide transcriptome analysis reveals Embryonic-Stem Cell derived tenocytes resemble fetal, not adult, tenocytes. Genome-wide transcriptome analysis reveals Embryonic-Stem Cell derived tenocytes resemble fetal, not adult, tenocytes

Purpose:- RNA sequencing was performed on adult, fetal and embryonic stem-cell derived equine tenocytes cultured in 3D to look at which had more similar gene expression profiles. Results:- 6,190 genes were significantly differentially expressed (DE) across the three lines of fetal, adult and ESC-tenocytes (logFC ± 2, padj <0.01). 542 of these were DE between adult and fetal tenocytes, 2,708 between fetal and ESC-tenocytes and 2,940 between adult and ESC-tenocytes. Genes which were DE between adult and fetal tenocytes were significanlty involved in cell migration and motility. Genes which were DE between the ESC-tenocytes and both adult and fetal tenocytes were in organism process and development. Overall design: Methods:- mRNA profiles of three independent lines of adult, fetal and ESC derived tenocytes were generated by RNA sequencing, using Illumina NovaSeq6000 and Illumina HiSeq2500. The sequence reads that passed quality filters were analyzed using the pseudoaligner Salmon and differential expression quantified using DESeq2, with batch correction applied. Gene ontology analysis was then conducted using Panther GoSlim.

研究目的：对体外三维培养的成年、胎儿及胚胎干细胞（ESC, Embryonic Stem Cell）来源的马肌腱细胞进行RNA测序，以探究不同来源肌腱细胞的基因表达谱相似性。研究结果：在胎儿、成年及胚胎干细胞来源的肌腱细胞三系细胞中，共鉴定出6190个显著差异表达（differentially expressed, DE）基因（logFC绝对值≥2，校正后P值<0.01）。其中，成年与胎儿肌腱细胞间存在542个差异表达基因，胎儿与胚胎干细胞来源肌腱细胞间存在2708个，成年与胚胎干细胞来源肌腱细胞间存在2940个。成年与胎儿肌腱细胞的差异表达基因显著富集于细胞迁移与运动相关生物学过程；胚胎干细胞来源肌腱细胞与成年、胎儿肌腱细胞的差异表达基因则显著富集于生物过程与发育相关通路。实验整体设计与方法：采用Illumina NovaSeq6000与Illumina HiSeq2500测序平台，对三株独立的成年、胎儿及胚胎干细胞来源的马肌腱细胞进行mRNA测序，获取其表达谱。对通过质量过滤的测序reads，使用伪比对工具Salmon进行序列比对分析，并通过DESeq2进行差异表达基因定量，同时应用批次校正方法。随后利用Panther GoSlim开展基因本体（Gene Ontology, GO）功能富集分析。