Department of Pathology and Laboratory Medicine, Weill Cornell Medicine, New York, NY, USA.
Research Division, Hospital for Special Surgery, New York, NY, USA.
J Bone Miner Res. 2019 Jul;34(7):1207-1219. doi: 10.1002/jbmr.3802.
Bone is composed of a complex mixture of many dynamic cell types. Flow cytometry and in vivo lineage tracing have offered early progress toward deconvoluting this heterogeneous mixture of cells into functionally well-defined populations suitable for further studies. Single-cell sequencing is poised as a key complementary technique to better understand the cellular basis of bone metabolism and development. However, single-cell sequencing approaches still have important limitations, including transcriptional effects of cell isolation and sparse sampling of the transcriptome, that must be considered during experimental design and analysis to harness the power of this approach. Accounting for these limitations requires a deep knowledge of the tissue under study. Therefore, with the emergence of accessible tools for conducting and analyzing single-cell RNA sequencing (scRNA-seq) experiments, bone biologists will be ideal leaders in the application of scRNA-seq to the skeleton. Here we provide an overview of the steps involved with a single-cell sequencing analysis of bone, focusing on practical considerations needed for a successful study. © 2019 American Society for Bone and Mineral Research.
骨骼由多种动态细胞类型组成的复杂混合物构成。流式细胞术和体内谱系追踪技术为将这种异质细胞混合物解析为适合进一步研究的功能明确的细胞群提供了早期进展。单细胞测序有望成为一种关键的补充技术，以更好地了解骨骼代谢和发育的细胞基础。然而，单细胞测序方法仍然存在重要的局限性，包括细胞分离的转录效应和转录组的稀疏采样，在实验设计和分析中必须考虑这些局限性，以利用该方法的优势。要克服这些局限性，需要深入了解研究中的组织。因此，随着单细胞 RNA 测序 (scRNA-seq) 实验的可及工具的出现，骨骼生物学家将成为 scRNA-seq 在骨骼中应用的理想领导者。本文提供了骨骼单细胞测序分析的步骤概述，重点介绍了成功进行研究所需的实际考虑因素。
