Current Chemical Genomics and Translational Medicine

2013, 7 : 21-29
Published online 2013 September 3. DOI: 10.2174/2213988501307010021
Publisher ID: CCGTM-7-21

RESEARCH ARTICLE
 High-Content Screening of Human Primary Muscle Satellite Cells for New Therapies for Muscular Atrophy/Dystrophy

Lidia S Nierobisz1 , Bentley Cheatham2 , Benjamin M Buehrer2 and Jonathan Z Sexton1, *
1 Biomanufacturing Research Institute and Technology Enterprise, Department of Pharmaceutical Sciences, North Carolina Central University Durham, NC 27707, USA
2 ZenBio Inc. RTP, NC 27709, USA

* Address correspondence to this author at the Biomanufacturing Research Institute and Technology Enterprise, North Carolina Central University, 1801 Fayetteville St., Durham, NC 27707, USA. Email: jsexton@nccu.edu

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.

Keywords: :

High-content screening, LY364947, muscle atrophy, phenotypic high-content analysis and satellite cell proliferation..