Bone Cell Biology and Disease Unit, St Vincent's Institute of Medical Research, 9 Princes St, Fitzroy, 3065, Australia.
Department of Medicine, The University of Melbourne, St. Vincent's Hospital, Melbourne, 3065, Australia.
Curr Osteoporos Rep. 2021 Dec;19(6):604-615. doi: 10.1007/s11914-021-00708-5. Epub 2021 Nov 10.
Osteocytes are cells embedded within the bone matrix, but their function and specific patterns of gene expression remain only partially defined; this is beginning to change with recent studies using transcriptomics. This unbiased approach can generate large amounts of data and is now being used to identify novel genes and signalling pathways within osteocytes both at baseline conditions and in response to stimuli. This review outlines the methods used to isolate cell populations containing osteocytes, and key recent transcriptomic studies that used osteocyte-containing preparations from bone tissue.
Three common methods are used to prepare samples to examine osteocyte gene expression: digestion followed by sorting, laser capture microscopy, and the isolation of cortical bone shafts. All these methods present challenges in interpreting the data generated. Genes previously not known to be expressed by osteocytes have been identified and variations in osteocyte gene expression have been reported with age, sex, anatomical location, mechanical loading, and defects in bone strength. A substantial proportion of newly identified transcripts in osteocytes remain functionally undefined but several have been cross-referenced with functional data. Future work and improved methods (e.g. scRNAseq) likely provide useful resources for the study of osteocytes and important new information on the identity and functions of this unique cell type within the skeleton.
骨细胞是嵌入骨基质中的细胞,但它们的功能和特定的基因表达模式仍未完全定义;随着最近使用转录组学的研究,这种情况开始发生变化。这种无偏方法可以生成大量数据,现在正被用于在基础条件和对刺激的反应中识别骨细胞内的新基因和信号通路。本综述概述了用于分离含有骨细胞的细胞群体的方法,以及最近使用骨组织中含有骨细胞的制剂进行的关键转录组学研究。
有三种常见的方法可用于准备样本以检查骨细胞的基因表达:消化后再进行排序、激光捕获显微镜和皮质骨轴的分离。所有这些方法在解释所产生的数据时都存在挑战。以前未知在骨细胞中表达的基因已被鉴定出来,并且随着年龄、性别、解剖位置、机械加载和骨强度缺陷的变化,骨细胞基因表达也发生了变化。在骨细胞中鉴定出的大量新转录本的功能仍然未定义,但其中一些已与功能数据交叉引用。未来的工作和改进的方法(例如 scRNAseq)可能为骨细胞的研究提供有用的资源,并为骨骼中这种独特细胞类型的特性和功能提供重要的新信息。