Transcriptome profiling reveals cell specific changes enabling osteogenic differentiation in response to mechanical stimulation

RNA-Seq was performed on 12 samples of cells grown in culture. There were two cell types, and three culture conditions. Two biological repeats were performed with each cell type in each condition. Human Dermal Papilla cells (DP) or human Papillary Fibroblasts (PFi) were grown in growth media (GM), Osteogenic media (OM), or Osteogenic media plus shockwave stimulation (OMSW). 24 hours after media change cells were homogenized through QIAshredders and total RNA was isolated using RNeasy Plus micro kit prior to library construction using SMART-seq2 prepartion. 75bp paired-end sequencing was carried out on Illumina HiSeq 4000 instrument.

本研究对12份体外培养的细胞样本开展了RNA测序（RNA-Seq）实验。实验设置2种细胞类型与3种培养条件，且每种细胞类型在每种培养条件下均设置2次生物学重复。所用细胞为人类真皮乳头细胞（Human Dermal Papilla cells, DP）或人类乳头成纤维细胞（human Papillary Fibroblasts, PFi），培养体系包括生长培养基（growth media, GM）、成骨培养基（Osteogenic media, OM）以及添加冲击波刺激的成骨培养基（Osteogenic media plus shockwave stimulation, OMSW）三种。更换培养基24小时后，使用QIAshredder柱对细胞进行匀浆处理，随后采用RNeasy Plus Micro试剂盒提取总RNA，再通过SMART-seq2方法完成文库构建。最终使用Illumina HiSeq 4000测序仪完成75bp双端测序。