Chapter 19 - High-Content Small Molecule Screening Strategies for Stem-Cell-Derived Motor Neurons

Human pluripotent stem cells possess great potential as a source of large numbers of defined cell populations that can be used for therapeutic applications and for pharmaceutical screening. By using disease-specific stem cells, in vitro disease models can be established that better recapitulate human diseases and lead to the development of more effective therapeutics and more informative disease mechanism studies. We have developed a high-content screening (HCS) assay using embryonic-stem-cell-derived motor neurons to study disease-related processes such as differentiation, survival, and protein expression. In particular, the assay described here is aimed at finding small molecules that can modulate the level of a key protein-survival of motor neuron-implicated in the human motor neuron disease-spinal muscular atrophy. A detailed protocol for the assay setups, including the parameters for growth conditions, compound library selection, and hit validation methods, are described.