High-throughput analysis of HGF-stimulated cell scattering.
Historically, only relatively low-throughput or expensive methods have been available to measure cell migration. Hepatocyte growth factor (HGF) is a ligand for the tyrosine kinase receptor Met that, in addition to mediating proliferation and survival, increases cell motility and metastasis. The authors have developed a high-throughput imaging assay for measuring inhibition of HGF-induced scattering in human HPAF-II pancreatic adenocarcinoma cells. Following treatment with test compounds and HGF for 24 h, cells are labeled with a nuclear stain and imaged at 10x magnification. The proximity of neighboring nuclei is measured, and the distribution of internuclear distances across each field of view is used to calculate the fraction of scattered cells. This method of analysis can be extended to other cell types and signaling pathways and, compared with other membrane-based migration assays currently available, the assay is significantly lower in cost, is less labor intensive, and provides higher throughput.
Chan
Grace K Y
GK
Department of Drug Design and Optimization/Automated Lead Optimization, Merck Research Laboratories Boston, Boston, Massachusetts 02115, USA.
Lutterbach
Bart A
BA
Pan
Bo-Sheng
BS
Kariv
Ilona
I
Szewczak
Alexander A
AA
D000230:Adenocarcinoma / Q000378:metabolism*; D001331:Automation; D045744:Cell Line, Tumor; D002465:Cell Movement; D002467:Cell Nucleus / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000378:metabolism*; D006801:Humans; D019044:Image Cytometry / Q000295:instrumentation* / Q000379:methods; D007091:Image Processing, Computer-Assisted; D020128:Inhibitory Concentration 50; D008024:Ligands; D008954:Models, Biological; D010190:Pancreatic Neoplasms / Q000378:metabolism*; D019859:Proto-Oncogene Proteins c-met / Q000378:metabolism
D008024:Ligands; D017228:Hepatocyte Growth Factor; D019859:Proto-Oncogene Proteins c-met