Advances in single-cell sequencing and its application to musculoskeletal system research

Abstract

In recent years, single-cell sequencing (SCS) technologies have continued to advance with improved operating procedures and reduced cost, leading to increasing practical adoption among researchers. These emerging technologies have superior abilities to analyse cell heterogeneity at a single-cell level, which have elevated multi-omics research to a higher level. In some fields of research, application of SCS has enabled many valuable discoveries, and musculoskeletal system offers typical examples. This article reviews some major scientific issues and recent advances in musculoskeletal system. In addition, combined with SCS technologies, the research of cell or tissue heterogeneity in limb development and various musculoskeletal system clinical diseases also provides new possibilities for treatment strategies. Finally, this article discusses the challenges and future development potential of SCS and recommends the direction of future applications of SCS to musculoskeletal medicine.

Keywords: cell heterogeneity; musculoskeletal system; single-cell multi-omics; single-cell sequencing.

FIGURE 1

An overview of the research issues of the musculoskeletal system

FIGURE 2

Basic procedures of single‐cell transcriptome sequencing (scRNA‐seq) and three common single‐cell sorting platform. ScRNA‐seq generally involves five basic procedures: single‐cell isolation, reverse transcription, cDNA amplification, library preparation and high‐throughput sequencing. Drop‐seq, 10X Genomics and BD Rhapsody are the most widely used single‐cell sorting platforms. They use microparticle beads with sequences linked by different cell barcodes, UMI and Oligo dT. These beads can capture mRNAs with poly A tails for reverse transcription and cDNA library preparation. Finally, all transcriptome information of a single cell is obtained by high‐throughput sequencing