Journal
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Volume 361, Issue 2, Pages 410-413Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2007.07.018
Keywords
monocyte; migration; chemotaxis; adhesion; fluorochrome; calcein AM; DMSO; growth factors; TGF-beta 1; fMLP
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For rapid chemotaxis quantification, cell prelabelling is often performed with the fluorochrorne calcein acetomethylester (calcein AM). We investigated whether calcein AM-prelabelling is reliable for monocyte migration analysis. Human monocytes were either pre-exposed to calcein AM or unlabelled. Monocyte migration towards the potent chemoattractants transforming growth factor-beta 1 (TGF-beta 1) and N-formyl-Methionin-Leucin-Phenylalanin (fMLP) was assessed using a 48-well micro-chemotaxis chamber. For quantification, cells were visualized by light microscopy and counted. Surprisingly, random migration of calcein AM-prelabelled cells was significantly impaired compared to the unlabelled control. Accordingly, monocyte chemotaxis towards either TGF-beta 1 or fMLP dramatically declined. Adherence of calcein AM-labelled monocytes on plastic was also significantly decreased compared to control cells. As adhesion is regarded as an essential component of monocyte migration, the reduced migration observed in calcein AM-labelled monocytes might be explained by a fluorochrome-induced adhesion defect. Therefore, use of the fluorochrome calcein AM cannot be recommended for functional testing of monocytes. (c) 2007 Elsevier Inc. All rights reserved.
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