Isolation, Purification, and Growth of Human Skeletal Muscle Cells

Skeletal muscle cells can be used in vitro for the study of myogenests, as well as in VIVO as gene-delivery vehicles for the therapy of muscle and nonmuscle diseases (1 –9 ). These skeletal muscle cells are derived from muscle satellite cells, which he between the basal lamina and the sarcolemma of differentiated muscle fibers (10 ) Normally quiescent after the period of muscle development and growth during fetal life and the early postnatal period, these cells are induced to proliferate on muscle damage and fuse with existing muscle fibers Satellite cells isolated and grown in vitro are called myoblasts Myoblasts proliferate in mitogen-rich media, but on reaching high cell density followed by exposure to mitogen-poor media, are induced to differentiate and become postmitotic Muscle differentiation is characterized by the fusion of myoblasts to form multinucleated myotubes, which express differentiation-specific proteins. In this chapter, methods are given for the isolation of myoblasts from human muscle tissue using two different techinques flow cytometry (11 ) and cell cloning (12 –14 ). These methods are applicable to muscle tissue from both fetal and postnatal donors, as well as from normal and diseased individuals