NDLI: Cell aggregation on agar as an indicator for cell-matrix adhesion: effects of opioids

Content Provider	Springer Nature Link
Author	Debruyne, Delphine Mareel, Marc Vanhoecke, Barbara Bracke, Marc
Copyright Year	2009
Abstract	The slow aggregation assay is generally used to study the functionality of cell–cell adhesion complexes. Single cells are seeded on a semisolid agar substrate in a 96-well plate and the cells spontaneously aggregate. We used HEK FLAG-MOP cells that stably overexpress the mu opioid receptor and the mu-opioid-receptor-selective agonists DAMGO and morphine to study whether other factors than functionality of cell–cell adhesions complexes can contribute to changes in the pattern of slow aggregation on agar. HEK FLAG-MOP cells formed small compact aggregates. In the presence of DAMGO and morphine, larger and fewer aggregates were formed in comparison to the vehicle control. These aggregates were localized in the center of the agar surface, whereas in the vehicle control they were dispersed over the substrate. However, in suspension culture on a Gyrotory shaker, no stimulation of aggregation was observed by DAMGO and morphine, showing that opioids do not affect affinity. A dissociation experiment revealed that HEK FLAG-MOP aggregates formed in the absence or presence of opioids are resistant to de-adhesion. We demonstrated that the larger aggregates are neither the result of cell growth stimulation by DAMGO and morphine. Since manipulations of the substrate such as increasing the agar concentration or mixing agar with agarose induced the same changes in the pattern of slow aggregation as treatment with opioids, we suggest that cell–substrate adhesion may be involved in opioid-stimulated aggregation.
Starting Page	473
Ending Page	482
Page Count	10
File Format	PDF
ISSN	10712690
Journal	In Vitro Cellular & Developmental Biology - Animal
Volume Number	45
Issue Number	8
e-ISSN	1543706X
Language	English
Publisher	Springer-Verlag
Publisher Date	2009-02-27
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Aggregation Agar Cell–cell adhesion Cell–substrate adhesion Opioids Animal Genetics and Genomics Cell Culture Stem Cells Developmental Biology Cell Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Developmental Biology Medicine

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