Quantitative assays of chemotaxis and chemokinesis for human neural cells.
Cell migration is vital for many physiological processes, and its modulation is likely to be of therapeutic benefit. In this study we have developed fluorescence image-based chemokinesis and chemotaxis assays on the Cellomics (Pittsburgh, PA) ArrayScan platform, which would be suitable for industrial drug discovery in a variety of fields. Studying the migratory characteristics of neural stem cells is of interest for understanding the therapeutic potential of these cells, in terms of both cellular transplantation and the activation of endogenous populations of stem cells. Growth conditions were identified whereby human neural precursors could be maintained as neurospheres and plated out into microtitre plates for high-content assays. Chemokinesis was assessed using fluorescent bead-coated 96-well microtitre plates, whilst chemotaxis was assessed using BD Biosciences (Oxford, UK) Fluoroblok 24-well plates. Assays for both chemokinesis and chemotaxis were developed that were quantified automatically using the ArrayScan and appropriate algorithms. Using the two complementary techniques, foetal bovine serum was observed to have chemokinetic effects on the cells, whilst platelet-derived growth factor isoform AB was chemotactic. The two assays described here are suitable for screening for novel modulators of cell migration, or for performing more detailed mechanistic follow-up studies. These assays enable us to perform cell motility studies with minimal laboratory handling, in an automated manner, thereby allowing quantitative studies of cell behaviour to be incorporated in a routine drug discovery screening cascade.
Richards
Gillian R
GR
The Neuroscience Research Centre, Merck Sharp and Dohme Research Laboratories, Terlings Park, Eastwick Road, Harlow, Essex CM20 2QR, UK.
Millard
Rachel M
RM
Leveridge
Mathew
M
Kerby
Julie
J
Simpson
Peter B
PB
D002452:Cell Count / Q000379:methods; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D002469:Cell Separation; D002478:Cells, Cultured; D018925:Chemokines / Q000502:physiology*; D002633:Chemotaxis / Q000187:drug effects / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology
D018925:Chemokines; D010982:Platelet-Derived Growth Factor