title: High-content screening of human primary muscle satellite cells for new therapies for muscular atrophy/dystrophy.
abstract: Myoblast proliferation and differentiation are essential for normal skeletal muscle growth and repair. Muscle recovery is dependent on the quiescent population of muscle stem cells - satellite cells. During muscle injury, satellite cells become mitotically active and begin the repair process by fusing with each other and/or with myofibers. Aging, prolonged inactivity, obesity, cachexia and other muscle wasting diseases are associated with a decreased number of quiescent and proliferating satellite cells, which impedes the repair process. A high-content/high-throughput platform was developed and utilized for robust phenotypic evaluation of human primary satellite cells in vitro for the discovery of chemical probes that may improve muscle recovery. A 1600 compound pilot screen was developed using two highly annotated small molecule libraries. This screen yielded 15 dose responsive compounds that increased proliferation rate in satellite cells derived from a single obese human donor. Two of these compounds remained dose responsive when counter-screened in 3-donor obese superlot. The Alk-5 inhibitor LY364947, was used as a positive control for assessing satellite cell proliferation/delayed differentiation. A multivariate approach was utilized for exploratory data analysis to discover proliferation vs. differentiation-dependent changes in cellular phenotype. Initial screening efforts successfully identified a number of phenotypic outcomes that are associated with desired effect of stimulation of proliferation and delayed differentiation.
authors.affiliation: Biomanufacturing Research Institute and Technology Enterprise, Department of Pharmaceutical Sciences, North Carolina Central University Durham, NC 27707, USA.
authors.forename: Lidia S
authors.initials: LS
authors.lastname: Nierobisz
authors.affiliation: ZenBio Inc. RTP, NC 27709, USA.
authors.forename: Bentley
authors.initials: B
authors.lastname: Cheatham
authors.affiliation: ZenBio Inc. RTP, NC 27709, USA.
authors.forename: Benjamin M
authors.initials: BM
authors.lastname: Buehrer
authors.affiliation: Biomanufacturing Research Institute and Technology Enterprise, Department of Pharmaceutical Sciences, North Carolina Central University Durham, NC 27707, USA.
authors.forename: Jonathan Z
authors.initials: JZ
authors.lastname: Sexton
keywords: High-content screening; LY364947; muscle atrophy; phenotypic high-content analysis and satellite cell proliferation.