Nitric Oxide as a Potential Adjuvant Therapeutic for Neuroblastoma: Effects of NO on Murine N2a Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. S-Nitrosoglutathione Synthesis
2.1.1. Materials
2.1.2. Synthesis of S-Nitrosoglutathione (GSNO)
2.2. Cell Culture—N2a
2.2.1. Materials
2.2.2. Method
2.3. Cell Culture—HDF
2.3.1. Materials
2.3.2. Method
2.4. In Vitro Cell Viability Assays
2.4.1. Materials
2.4.2. Method
2.4.3. Resazurin Assay
2.4.4. WST-8 Assay
2.4.5. MTT Assay
2.5. Colony Formation
Method
2.6. Nitric Oxide Analyzer Analysis of 1 mM GSNO in PBS
2.7. UV-Vis Analysis of 1 mM GSNO in Complete DMEM
2.8. Data Analysis/Statistics
3. Results
3.1. Cell Viability Assays
3.1.1. Resazurin Assay—Analysis of Cellular Viability
3.1.2. WST-8 Assay—Analysis of Cellular Viability
3.1.3. MTT Assay–Analysis of Cellular Viability
3.2. Colony Formation Assay
3.3. Impact of NO on HDFa Cells—Resazurin Assay
3.4. NO-Release from 1 mM GSNO
3.4.1. NO-Release from 1 mM GSNO in PBS
3.4.2. Absorbance of GSNO λmax via UV/VIS
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Time (h) | Absorbance at 336 nm (GSNO λmax) | Concentration of GSNO (mM) | Amount of Available NO (μmol) |
---|---|---|---|
0 h | 0.895 | 0.971 | 0.433 |
6 h | 0.598 | 0.649 | 0.290 |
12 h | 0.304 | 0.330 | 0.147 |
18 h | 0.130 | 0.141 | 0.0630 |
24 h | 0.135 | 0.146 | 0.0650 |
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Gordon, J.L.; Reynolds, M.M.; Brown, M.A. Nitric Oxide as a Potential Adjuvant Therapeutic for Neuroblastoma: Effects of NO on Murine N2a Cells. Vet. Sci. 2020, 7, 51. https://doi.org/10.3390/vetsci7020051
Gordon JL, Reynolds MM, Brown MA. Nitric Oxide as a Potential Adjuvant Therapeutic for Neuroblastoma: Effects of NO on Murine N2a Cells. Veterinary Sciences. 2020; 7(2):51. https://doi.org/10.3390/vetsci7020051
Chicago/Turabian StyleGordon, Jenna L., Melissa M. Reynolds, and Mark A. Brown. 2020. "Nitric Oxide as a Potential Adjuvant Therapeutic for Neuroblastoma: Effects of NO on Murine N2a Cells" Veterinary Sciences 7, no. 2: 51. https://doi.org/10.3390/vetsci7020051