The edge effect: A global problem. The trouble with culturing cells in 96-well plates.
96-Well plates
Cell culture
Edge effect
Journal
Biochemistry and biophysics reports
ISSN: 2405-5808
Titre abrégé: Biochem Biophys Rep
Pays: Netherlands
ID NLM: 101660999
Informations de publication
Date de publication:
Jul 2021
Jul 2021
Historique:
received:
23
05
2020
revised:
15
03
2021
accepted:
16
03
2021
entrez:
15
4
2021
pubmed:
16
4
2021
medline:
16
4
2021
Statut:
epublish
Résumé
The use of 96-well plates is ubiquitous in preclinical studies. Corner and edge wells have been observed to be more prone to evaporation compared to interior wells. Mammalian cells were cultured in 96-well plates over a period of 72 h. VWR and Greiner plates were tested. MTS reagent was added, and metabolic activity was determined after 2 h. When using VWR plates, cells showed a highly heterogeneous pattern of cell growth. The outer wells showed 35% lower metabolic activity than the central wells. Cells grown in rows two and three also grew sub-optimally (25% and 10% reduction compared to central wells). Greiner plates showed better homogeneity. Cells grown in the outer wells showed 16% lower metabolic activity while cells in rows two and three showed reductions of 7 and 1%, respectively. This edge effect was partially mitigated by storing the plates in loosely sealed wrapping during incubation. Placing a buffer between the wells of the plate further improved homogeneity for the Greiner plates. Different brands of 96-well plates show different levels of the edge effect. Some clearly are inappropriate for such studies. Each laboratory needs to determine their own optimum conditions for culturing cells empirically before continuing to use multiwell plates. Otherwise, large artifacts may arise, affecting the quality of data, with the potential of introducing type I or type II errors.
Sections du résumé
BACKGROUND
BACKGROUND
The use of 96-well plates is ubiquitous in preclinical studies. Corner and edge wells have been observed to be more prone to evaporation compared to interior wells.
METHODS
METHODS
Mammalian cells were cultured in 96-well plates over a period of 72 h. VWR and Greiner plates were tested. MTS reagent was added, and metabolic activity was determined after 2 h.
RESULTS
RESULTS
When using VWR plates, cells showed a highly heterogeneous pattern of cell growth. The outer wells showed 35% lower metabolic activity than the central wells. Cells grown in rows two and three also grew sub-optimally (25% and 10% reduction compared to central wells). Greiner plates showed better homogeneity. Cells grown in the outer wells showed 16% lower metabolic activity while cells in rows two and three showed reductions of 7 and 1%, respectively. This edge effect was partially mitigated by storing the plates in loosely sealed wrapping during incubation. Placing a buffer between the wells of the plate further improved homogeneity for the Greiner plates.
CONCLUSION
CONCLUSIONS
Different brands of 96-well plates show different levels of the edge effect. Some clearly are inappropriate for such studies.
GENERAL SIGNIFICANCE
CONCLUSIONS
Each laboratory needs to determine their own optimum conditions for culturing cells empirically before continuing to use multiwell plates. Otherwise, large artifacts may arise, affecting the quality of data, with the potential of introducing type I or type II errors.
Identifiants
pubmed: 33855228
doi: 10.1016/j.bbrep.2021.100987
pii: S2405-5808(21)00081-9
pmc: PMC8024881
doi:
Types de publication
Journal Article
Langues
eng
Pagination
100987Informations de copyright
© 2021 The Authors. Published by Elsevier B.V.
Déclaration de conflit d'intérêts
This research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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