Spin infection enables efficient gene delivery to muscle stem cells. (August 2017)

• Record Type: Journal Article
• Authors: Kodaka, Yusaku; Asakura, Yoko; Asakura, Atsushi
• Is Part Of: Biotechniques. Volume 63:Number 2(2017)
• Page Start: 72
• Page End: 76
• Publication Date: 2017-08
• Subjects: muscle stem cell -- satellite cell -- skeletal muscle -- regeneration -- retroviral vector; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000114576
• Languages: English
• ISSNs: 0736-6205

Abstract

Viral vector–mediated foreign gene expression in cultured cells has been extensively used in stem cell studies to explore gene function. However, it is difficult to obtain high-quality stem cells and primary cells after viral vector infection. Here, we describe a new protocol for high-efficiency retroviral infection of primary muscle stem cell (satellite cell) cultures. We compared multiple commercially available transfection reagents to determine which was optimal for retroviral infections of primary myoblasts. Centrifugation force was also tested, and a spin infection protocol with centrifugation at 2800 × g for 90 min had the highest infection efficiency for primary myoblasts. We confirmed that infected muscle stem cells maintain cell proliferation and the capacity for in vitro and in vivo myogenic differentiation. Our new, efficient retroviral infection protocol for muscle stem cells can be applied to molecular biology experiments as well as translational studies.