Abstract: Single-cell RNA sequencing (scRNA-seq) technology is used to sequence the transcriptome at the single cell level,which can describe samples from multiple perspectives such as expression amount,cell size and cell composition,mainly including single cell isolation,mRNA reverse transcription,library construction,transcriptome sequencing and data analysis.Single cell isolation is the first step of this technology. Common cell isolation methods include serial dilution,micromanipulation,fluorescent activated cell sorting,laser capture microdissection and microfluidics.Many transcriptome sequencing methods based on different cell capture,cDNA amplification and library construction have been developed,such as CEL-seq2,Drop-seq,MARS-seq,SCRB-seq,Smart-seq,Smart-seq2,etc.Compared with the traditional sequencing method,scRNA-seq can identify the gene expression information of a single cell,record the spatial location of the cell,and track the trajectory of different cell lineages in the differentiation process,which brings great convenience for studying the molecular specificity of intramuscular adipocytes that are difficult to extract in large quantities and the occurrence and differentiation process.The author briefly introduced the key steps of scRNA-seq technology,compared and analyzed the advantages and disadvantages of single cell isolation and transcriptome sequencing methods,reviewed the origin of intramuscular adipocytes,and the application of scRNA-seq in the identification of subgroups and marker genes of intramuscular adipocytes,as well as the tracking of cell differentiation trajectory,in order to provide a reference for the further study of intramuscular adipocytes.

Key words: single-cell RNA sequencing; intramuscular adipocytes; cell heterogeneity