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Cell passaging rapidly affects expression, secretion and
activity of MMP9 as well as mobility of HL60 leukemia cells
Yohann Bernard, Sébastien Plançon,
Chantal Melchior, Eric Tschirhart and Jean-Luc Bueb*
Université du Luxembourg, Life Sciences Research Unit, 162a,
avenue de la Faïencerie, L-1511 Luxembourg, Luxembourg.
*Corresponding author. E-mail: jean-luc.bueb@uni.lu. Tel :
(+352) 466 644 6738. Fax :(+352) 466 644 6435.
Accepted 6th August, 2008 |
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The HL60 cell line, derived from acute promyelocytic
leukemia cells, can differentiate into neutrophil-like cell
following DMSO treatment. Mobility of HL60, or
DMSO-differentiated HL60 cells (≠HL60), requires surface
expression of adhesion molecules and production of matrix
metalloproteinases (MMPs). The aim of this study was to
investigate in HL60 and ≠HL60 the effects of cell passaging
(over 5 passages after delivery (P and P+5)) on i) surface
expression of adhesion molecule CD11b, which is considered a
neutrophil differentiation marker ii) MMP9 mRNA expression,
protein release and zymographic activity and iii) cellular
mobility. As expected, CD11b expression at both cell
passages increased in ≠HL60 relative to undifferentiated
HL60, but expression levels of this neutrophils marker did
not change over 5 passages. MMP9 mRNA expression however, in
basal conditions was increased in HL60 at P+5. At P+5 versus
P, MMP9 protein levels, MMP9 zymographic activity and
cellular mobility in HL60 and ≠HL60 were elevated.
Stimulation by N-formyl-L-Methionyl-L-Leucyl-L-Phenylalanine
had no effects on HL60, but raised MMP9 protein
concentration and zymographic activity in ≠HL60. Since
passage history is likely to also influence cellular
functions other than MMP-related effects, it is important to
carefully consider passage numbers when designing
experiments.
Key words: Matrix metalloproteinases, mobility, cell
passaging, HL60 cell line, DMSO-differentiation |
