Methods Mol Biol. 2019;1889:81-93. doi: 10.1007/978-1-4939-8897-6_6.

Isolation and Purification of Satellite Cells from Young Rats by Percoll Density Gradient Centrifugation.

Matsuyoshi Y¹, Akahoshi M¹, Nakamura M¹, Tatsumi R¹, Mizunoya W².

Abstract

Satellite cells (SCs) are myogenic stem cells that play an important role in skeletal muscle regeneration and hypertrophy. Primary cultures of SCs are useful to analyze cell functions; however, it is difficult to obtain highly pure SCs from young rats with the conventional procedures. The purpose of this study is to establish a purification method for SC isolation from young rats and quantitatively evaluate the purification procedure employing Percoll, a common research tool to purify cells. We elucidated the purity of SCs collected by Percoll density gradient centrifugation using real-time RT-qPCR and immunocytochemistry for desmin. Percoll treatment increased the purity of SCs isolated from young rats to nearly 90%, which was comparable to that achieved with the conventional method using middle-aged rats.