High-Content Screening of Human Primary Muscle Satellite Cells for New Therapies for Muscular Atrophy/Dystrophy
Results are encouraging
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 com- pounds that increased proliferation rate in satellite cells derived from a single obese human donor. Two of these com- pounds 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.
We developed a novel high content assay to detect stimulated proliferation of primary human skeletal muscle satellite cells. This scatterplot shows the results for 1600 compounds screened, including all FDA-approved drugs and 500 natural products. We discovered that 15 compounds in this set were capable of stimulating satellite cells to induce proliferation. The cells used during this screen were obtained as a tissue sample of rectus abdominis from consensual donors.
A positive result
In this screen, we found three very interesting natural classes of compounds; proliferators, toxic compounds and myogenic compounds. This becomes apparent when we plot proliferative status (cell number) versus cell size.
The most interesting compounds from this screen were Geraldol and Bromopride that consistently increased proliferation rates in satellite cells.