A morphology- and kinetics-based cascade for human neural cell high content screening.
The prospect of manipulating endogenous neural stem cells to replace damaged tissue and correct functional deficits represents a novel mechanism for treating a variety of central nervous system disorders. Using human neural precursor cultures and a variety of assays for studying stem cell behavior we have screened two libraries of commercially available compounds using an endpoint high content screening assay. We then performed detailed follow-up mechanistic studies on confirmed hits using endpoint and kinetics assays to characterize and differentiate the mechanisms of action of these compounds. The screening cascade employed successfully identified a number of active compounds with differing mechanisms of action. This approach shows how hits from a phenotypic screen can be prioritized and characterized by high content screening to identify potentially novel mechanisms and druggable targets to take forward into more conventional high-throughput screening approaches.
Richards
Gillian R
GR
The Neuroscience Research Centre, Merck Sharp and Dohme Research Laboratories, Harlow, Essex, UK. gillian_richards@merck.com
Smith
Alison J
AJ
Parry
Frances
F
Platts
Amy
A
Chan
Grace K Y
GK
Leveridge
Mathew
M
Kerby
Julie E
JE
Simpson
Peter B
PB
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical; D005260:Female; D006801:Humans; D007700:Kinetics; D016501:Neurites / Q000502:physiology / Q000648:ultrastructure; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D051381:Rats; D015398:Signal Transduction / Q000502:physiology; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology
D002118:Calcium