Abstract
Skeletal muscle from meat-producing livestock such as cattle is a major source of food for humans. To improve skeletal muscle growth efficiency or quality in cattle, it is necessary to understand the genetic and physiological mechanisms that govern skeletal muscle composition, development, and growth. Satellite cells are the myogenic progenitor cells in postnatal skeletal muscle. In this study we analyzed the composition of bovine satellite cells with single-cell RNA sequencing (scRNA-seq). We isolated satellite cells from a 2-week-old male calf, cultured them in growth medium for a week, and performed scRNA-seq using the 10x Genomics platform. Deep sequencing of two scRNA-seq libraries constructed from cultured bovine satellite cells yielded 860 million reads. Cell calling analyses revealed that these reads were sequenced from 19,096 individual cells. Clustering analyses indicated that these reads represented 15 cell clusters that differed in gene expression profile. Based on the enriched expression of markers of satellite cells (PAX7 and PAX3), markers of myoblasts (MYOD1, MYF5), and markers of differentiated myoblasts or myocytes (MYOG), three clusters were determined to be satellite cells, two clusters myoblasts, and two clusters myocytes. Gene ontology and trajectory inference analyses indicated that cells in these myogenic clusters differed in proliferation rate and differentiation stage. Two of the remaining clusters were enriched with PDGFRA, a marker of fibro-adipogenic (FAP) cells, the progenitor cells for intramuscular fat, and are therefore considered to be FAP cells. Gene ontology analyses indicated active lipogenesis in one of these two clusters. The identity of the remaining six clusters could not be defined. Overall, the results of this study support the hypothesis that bovine satellite cells are composed of subpopulations that differ in transcriptional and myogenic state. The results of this study also support the hypothesis that intramuscular fat in cattle originates from fibro-adipogenic cells.
Highlights
IntroductionScRNA-seq has proved to be a powerful method for assessing the heterogeneity of a cell population and for identifying rare or previously uncharacterized cell types in complex organs and tissues (Tang et al, 2009; Andrews and Hemberg, 2018)
Single-cell RNA sequencing is a relatively new technology that analyzes transcriptomes in individual cells by deep sequencing (Tang et al, 2009). scRNA-seq has proved to be a powerful method for assessing the heterogeneity of a cell population and for identifying rare or previously uncharacterized cell types in complex organs and tissues (Tang et al, 2009; Andrews and Hemberg, 2018).Satellite cells are myogenic progenitor cells in skeletal muscle of postnatal animals (Mauro, 1961)
Cultured Bovine Satellite Cells Are Heterogenous in Gene Expression
Summary
ScRNA-seq has proved to be a powerful method for assessing the heterogeneity of a cell population and for identifying rare or previously uncharacterized cell types in complex organs and tissues (Tang et al, 2009; Andrews and Hemberg, 2018). Satellite cells are myogenic progenitor cells in skeletal muscle of postnatal animals (Mauro, 1961). Satellite cells play an essential role in skeletal muscle regeneration and growth (Dhawan and Rando, 2005). Satellite cells are normally quiescent but are activated under conditions such as muscle injury (Dhawan and Rando, 2005). Satellite cells proliferate as myoblasts, differentiate, and fuse with each other to generate muscle fibers or with existing muscle fibers to increase muscle size
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