Improved agarose gel assay for quantification of growth factor-induced cell motility.
Cell chemotaxis is frequently required in normal or pathological situations such as invasion, metastasis, and tumor angiogenesis and may involve many different cell types. At present, no device can simultaneously (i) make morphological observations, (ii) quantify cell migration, (iii) test multiple chemoattracting gradients, and (iv) analyze cell-cell interactions. We developed an agarose-based assay to address these questions. Two glass molds were designed, around which agarose gel could be poured to form specific well shapes. Using a vital nuclear stain (Hoechst 33258), we characterized the migration profile of adherent or suspension cells. Cells could be observed during the entire migration process. We were able to follow cells moving toward chemoattractants or being repulsed by other molecules, and we could estimate average migration speed. Using this inexpensive assay, we were able to obtain precise, reproducible results concerning the chemotactic behavior of different cell types. The resulting data differentiated between chemokinetic and chemotactic movement. Chemotactic potencies could be compared using different criteria, such as the number of attracted cells, induced speed, and morphological aspect. This improved agarose assay appears to be a reliable and inexpensive alternative to other available chemotaxis study tools.
Mousseau
Yoanne
Y
University of Limoges, Limoges, France.
Leclers
Dany
D
Faucher-Durand
Karine
K
Cook-Moreau
Jeanne
J
Lia-Baldini
Anne-Sophie
AS
Rigaud
Michel
M
Sturtz
Franck G
FG
D001681:Biological Assay / Q000295:instrumentation* / Q000379:methods; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002633:Chemotaxis / Q000187:drug effects* / Q000502:physiology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods; D017022:Flow Injection Analysis / Q000295:instrumentation* / Q000379:methods; D005782:Gels / Q000737:chemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000008:administration & dosage*; D012685:Sepharose / Q000737:chemistry*
D005782:Gels; D036341:Intercellular Signaling Peptides and Proteins; D012685:Sepharose