Efficient Transduction and Expansion of Ovine Macrophages for Gene Therapy Implementations
Abstract
:1. Introduction
2. Materials and Methods
2.1. Isolation of Ovine Monocytes
2.2. Macrophage Phagocytosis Assay
2.3. Lentiviral Vector Production and Titration
2.4. Macrophage Transduction
2.5. Vector Copy Number Analysis
2.6. Statistics
3. Results
3.1. Characterization of Peripheral Blood-Derived Ovine Macrophages
3.2. Phagocytosis Assay
3.3. Ovine Macrophages’ Resistance to Transduction May Be Averted with Transduction Enhancers
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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1st Experiment | 2nd Experiment | 3rd Experiment | |
---|---|---|---|
Control | 0.1 | 0.17 | 0.13 |
0.2 μm | 1.15 | 1.21 | 1 |
0.5 μm | 1.7 | 2.6 | 2.2 |
1 μm | 3.6 | 6 | 5.75 |
2 μm | 8.9 | 8.6 | 10 |
1st Experiment | 2nd Experiment | 3rd Experiment | 4th Experiment | 5th Experiment | ||
---|---|---|---|---|---|---|
No additive | control | 0.27 | 0.09 | 1.4 | 0.9 | 1.54 |
MOI *:60 | 48.7 | 48.1 | 43.7 | 34.2 | 18.3 | |
MOI:6 | 1.67 | 1.66 | 8.8 | 2.7 | 2.37 | |
MOI:0.6 | 0.35 | 0.16 | 1.7 | 1.9 | 1.47 | |
Polybrene | control | 0.27 | 0.09 | 0.86 | 0.7 | 0.95 |
MOI:60 | 97.34 | 97.46 | 78 | 75.6 | 51.4 | |
MOI:6 | 45.9 | 42.7 | 14.55 | 14.7 | 2.75 | |
MOI:0.6 | 1.5 | 0.71 | 1.34 | 1.25 | 0.91 | |
Protamine sulfate | control | 0.3 | 0.15 | 1.04 | 1.05 | 1.14 |
MOI:60 | 97.5 | 94.7 | 70 | 64.7 | 40.3 | |
MOI:6 | 21.2 | 16.8 | 6.8 | 10.8 | 7.55 | |
MOI:0.6 | 0.54 | 0.41 | 1.06 | 2.8 | 1.76 |
Days Post Transduction | MOI: 60 | |
---|---|---|
No additive | 3 | 10.46 |
8 | 0.13 | |
Polybrene | 3 | 11.33 |
8 | 0.29 | |
Protamine sulfate | 3 | 10.94 |
8 | 0.21 |
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Karponi, G.; Kritas, S.; Petridou, E.; Papanikolaou, E. Efficient Transduction and Expansion of Ovine Macrophages for Gene Therapy Implementations. Vet. Sci. 2018, 5, 57. https://doi.org/10.3390/vetsci5020057
Karponi G, Kritas S, Petridou E, Papanikolaou E. Efficient Transduction and Expansion of Ovine Macrophages for Gene Therapy Implementations. Veterinary Sciences. 2018; 5(2):57. https://doi.org/10.3390/vetsci5020057
Chicago/Turabian StyleKarponi, Garyfalia, Spyridon Kritas, Evanthia Petridou, and Eleni Papanikolaou. 2018. "Efficient Transduction and Expansion of Ovine Macrophages for Gene Therapy Implementations" Veterinary Sciences 5, no. 2: 57. https://doi.org/10.3390/vetsci5020057