Abstract
The objective of this study was to establish the optimum protocol for the isolation and culture of porcine muscle satellite cells. Mononuclear muscle satellite cells are a kind of adult stem cell, which is located between the basal lamina and sarcolemma of muscle fibers and is the primary source of myogenic precursor cells in postnatal muscle. Muscle satellite cells are a useful model to investigate the mechanisms of muscle growth and development. Although the isolation and culture protocols of muscle satellite cells in some species (e.g. mouse) have been established successfully, the culture system for porcine muscle satellite cells is very limited. In this study, we optimized the isolation procedure of porcine muscle satellite cells and elaborated the isolation and culture process in detail. Furthermore, we characterized the porcine muscle satellite cells using the immunofluorecence. Our study provides a reference for the isolation of porcine muscle satellite cells and will be useful for studying the molecular mechanisms in these cells.
Highlights
In livestock animals, skeletal muscle is the main component of lean body mass and its growth is the major factor affecting body growth
Detailed isolation procedures for porcine skeletal muscle satellite cells were not described in these studies, and these methods were time-consuming due to cumbersome steps and produced a low number of satellite cells, which limited the studies on muscle growth and development in pigs
The freezing medium was prepared with dimethyl sulfoxide (DMSO) and fetal bovine serum (FBS) according to the ratio of 1:5, e.g. 1 mL freezing medium was prepared with 200 μL DMSO and 800 μL FBS, and gently mixed
Summary
Skeletal muscle is the main component of lean body mass and its growth is the major factor affecting body growth. Porcine skeletal muscle satellite cells were first isolated and cultured in 1992 (Doumit and Merkel, 1992). Afterwards, porcine skeletal muscle satellite cells were isolated using similar or improved methods and were used to investigate the mechanisms of skeletal muscle growth and development (Mesires and Doumit, 2002; Theil et al, 2006; Mau et al, 2008; Wilschut et al, 2010). Detailed isolation procedures for porcine skeletal muscle satellite cells were not described in these studies, and these methods were time-consuming due to cumbersome steps and produced a low number of satellite cells, which limited the studies on muscle growth and development in pigs. The isolating processes for porcine skeletal muscle satellite cells were optimized and elaborated in detail, and the characterization of the isolated porcine skeletal muscle
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