Institute for Medical Genetics and Human Genetics, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Berlin, Germany; Department of Biomedical Engineering, Faculty of Electrical Engineering and Communication, Brno University of Technology, Brno, Czech Republic.
Institute for Medical Genetics and Human Genetics, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Berlin, Germany.
Bone. 2018 Aug;113:29-40. doi: 10.1016/j.bone.2018.04.006. Epub 2018 Apr 11.
Osteoblastic differentiation is a multistep process characterized by osteogenic induction of mesenchymal stem cells, which then differentiate into proliferative pre-osteoblasts that produce copious amounts of extracellular matrix, followed by stiffening of the extracellular matrix, and matrix mineralization by hydroxylapatite deposition. Although these processes have been well characterized biologically, a detailed transcriptional analysis of murine primary calvaria osteoblast differentiation based on RNA sequencing (RNA-seq) analyses has not previously been reported. Here, we used RNA-seq to obtain expression values of 29,148 genes at four time points as murine primary calvaria osteoblasts differentiate in vitro until onset of mineralization was clearly detectable by microscopic inspection. Expression of marker genes confirmed osteogenic differentiation. We explored differential expression of 1386 protein-coding genes using unsupervised clustering and GO analyses. 100 differentially expressed lncRNAs were investigated by co-expression with protein-coding genes that are localized within the same topologically associated domain. Additionally, we monitored expression of 237 genes that are silent or active at distinct time points and compared differential exon usage. Our data represent an in-depth profiling of murine primary calvaria osteoblast differentiation by RNA-seq and contribute to our understanding of genetic regulation of this key process in osteoblast biology.
成骨细胞分化是一个多步骤的过程，其特征是间充质干细胞的成骨诱导，然后分化为增殖性成骨前体细胞，产生大量细胞外基质，随后细胞外基质变硬，通过羟基磷灰石沉积进行基质矿化。尽管这些过程在生物学上已经得到很好的描述，但以前没有基于 RNA 测序 (RNA-seq) 分析对鼠原代颅骨成骨细胞分化进行详细的转录组分析。在这里，我们使用 RNA-seq 在体外分化的四个时间点获得了 29148 个基因的表达值，直到通过显微镜检查清楚地检测到矿化开始。标记基因的表达证实了成骨分化。我们使用无监督聚类和 GO 分析探索了 1386 个蛋白编码基因的差异表达。通过与位于同一拓扑关联域内的蛋白编码基因的共表达研究了 100 个差异表达的 lncRNA。此外，我们监测了 237 个在不同时间点沉默或活跃的基因的表达，并比较了差异外显子的使用。我们的数据通过 RNA-seq 对鼠原代颅骨成骨细胞分化进行了深入分析，有助于我们理解成骨细胞生物学中这一关键过程的遗传调控。
