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A new cytotoxicity assay for brevetoxins using fluorescence microscopy
Jennifer R McCall1, Elizabeth A Elliott2, Andrea J Bourdelais3
1UNCW Center for Marine Science, 5600 Marvin K Moss Lane, Wilmington, NC 28409, USA. mccalljr@uncw.edu.
Marine Drugs
|September 25, 2014
Summary
Brevetoxin detection assays using Neuro-2A cells show variability. A new fluorescence imaging model offers accurate brevetoxin cytotoxicity assessment and identifies SJCRH30 cells as a promising alternative without pre-treatment.
Area of Science:
- Marine toxicology
- Cellular toxicology
- Biomarker discovery
Background:
- Brevetoxins, produced by Karenia brevis, cause neurotoxic shellfish poisoning via sodium channel activation.
- Current brevetoxin detection relies on Neuro-2A cells requiring ouabain/veratridine pre-treatment.
- The traditional assay exhibits EC50 variability and non-sigmoidal dose-response curves.
Purpose of the Study:
- To address limitations in brevetoxin cytotoxicity assays.
- To develop a more accurate and reliable method for brevetoxin detection.
- To evaluate alternative cell lines for brevetoxin testing.
Main Methods:
- Development of a novel fluorescence live cell imaging model for cytotoxicity assessment.
- Utilized nuclear staining and other viability parameters for accurate measurement.
- Compared the performance of the new model and SJCRH30 cells against the traditional Neuro-2A assay.
Main Results:
- The traditional Neuro-2A assay demonstrates significant variability in EC50 values and non-linear dose-response curves.
- The new fluorescence live cell imaging model enables accurate cytotoxicity calculation.
- SJCRH30 cells show potential as an alternative cell line, negating the need for ouabain and veratridine pre-treatment.
Conclusions:
- The traditional brevetoxin assay using Neuro-2A cells is unreliable due to inherent variability.
- A new fluorescence live cell imaging assay provides a more accurate method for brevetoxin detection.
- SJCRH30 cells represent a viable alternative for brevetoxin testing, simplifying the assay process.
