Detecting Neurodevelopmental Toxicity of Domoic Acid and Ochratoxin A Using Rat Fetal Neural Stem Cells
Abstract
:1. Introduction
2. Results
2.1. rNSC Proliferation Without Differentiation
2.2. Neurosphere Assay
2.2.1. Neurosphere Proliferation Without Differentiation
2.2.2. Neurosphere Differentiation into Oligodendrocytes
2.3. rNSC Monolayer-Based Models for Three Types of Differentiation Processes
2.3.1. rNSC Differentiation Directed into Oligodendrocytes
2.3.2. rNSC Differentiation Directed into Astrocytes
2.3.3. rNSC Differentiation Directed into Neurons
2.3.4. Effects of Domoic Acid on rNSC Differentiation Directed into Astrocytes, Neurons, and Oligodendrocytes
2.4. Effects of DA and OTA on Cytotoxicity and on the Differentiation of rNSC Directed into Astrocytes
2.5. Effects of DA and OTA on Cytotoxicity and on the Differentiation of rNSC Directed into Neurons
2.6. Effects of DA and OTA on Cytotoxicity and on the Differentiation of rNSC Directed into Oligodendrocytes
2.7. Effects of DA and OTA on the Axonal Length of the Mature Neurons
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Culture
4.2.1. Propagation of rNSC
4.2.2. rNSC Neurosphere-Based Model for Differentiation
4.2.3. rNSC Monolayer-Based Model for Differentiation
A. Coating Chamber Slides for Differentiation into Various Cell Types
B. Differentiating Media
C. rNSC Culturing for Differentiation
4.3. Cytotoxicity Assays (Relative Cell Count)
4.4. Effects of DA and OTA on Directed Differentiation on the Different Cell Types
4.5. Immunochemistry
4.6. Image Analysis
4.7. Tracing and Measurement of the Axonal Length of the Mature Neurons
4.8. Statistical Analysis
5. Conclusion and Future Perspectives
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
BBB | Blood Brain Barrier |
CNS | Central Nervous System |
DA | Domoic Acid |
DNT | Developmental Neurotoxicology |
EGF | Epidermal Growth Factor |
FGFb | Fibroblast Growth Factor basic |
GFAP | Glial Fibrillary Acidic Protein |
MAP2 | Microtubule-Associated Protein 2 |
OTA | Ochratoxin A |
PSC | Pluripotent Stem Cells |
rNSC | rat fetal Neural Stem Cells |
RCC | Relative Cell Count |
RT | Room Temperature |
T3 | Thyroxin hormone |
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Gill, S.; Kumara, V.M.R. Detecting Neurodevelopmental Toxicity of Domoic Acid and Ochratoxin A Using Rat Fetal Neural Stem Cells. Mar. Drugs 2019, 17, 566. https://doi.org/10.3390/md17100566
Gill S, Kumara VMR. Detecting Neurodevelopmental Toxicity of Domoic Acid and Ochratoxin A Using Rat Fetal Neural Stem Cells. Marine Drugs. 2019; 17(10):566. https://doi.org/10.3390/md17100566
Chicago/Turabian StyleGill, Santokh, and V. M. Ruvin Kumara. 2019. "Detecting Neurodevelopmental Toxicity of Domoic Acid and Ochratoxin A Using Rat Fetal Neural Stem Cells" Marine Drugs 17, no. 10: 566. https://doi.org/10.3390/md17100566