Optimization of the Amplification of Equine Muscle-Derived Mesenchymal Stromal Cells in a Hollow-Fiber Bioreactor
Abstract
:1. Introduction
2. Material and Methods
2.1. Cell Cultures
2.1.1. Equine Muscle Microbiopsy and Mesenchymal Stromal Cell Culture
2.1.2. Quantum™ Preparation and Amplification of the Equine Muscle-Derived Mesenchymal Stromal Cells
Priming and Coating
Loading and Feeding
Cell Harvest
2.2. Flow Cytometry Analysis of the Immunophenotype of the Generated Cells
2.3. Trilineage Differentiation of Generated Equine Mesenchymal Stromal Cells
3. Results
Muscle-Derived Mesenchymal Stromal Cell Culture, Amplification and Characterization
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Antibody | Clone | Dilution |
---|---|---|
CD-44 | CVS18 | 25 |
CD-45 | F-10-89-4 | 5 |
MHCII | CVS20 | 25 |
CD-90 | DH24A | 50 |
Run | Horse | Passage | Fresh/ Frozen | Loading | Seeding | Duration (Days) | Harvest | Cell Viability | Final Count (Viable Cells) | Multiplication Factor |
---|---|---|---|---|---|---|---|---|---|---|
1 | Horse 1 | P5 | Fresh | Classic | 25 × 106 | 7 | 217 × 106 | 91% | 197 × 106 | 7.88 |
2 | Horse 1 | P3 | Fresh | Classic | 10 × 106 | 15 | 30 × 106 | 60% | 18 × 106 | 1.80 |
3 | Horse 1 | P6 | Frozen | Classic | 16.5 × 106 | 8 | 79 × 106 | 73% | 57.7 × 106 | 3.50 |
4 | Horse 2 | P6 | Fresh | Classic | 25 × 106 | 8 | 92 × 106 | 78% | 71.8 × 106 | 2.87 |
5 | Horse 3 | P4 | Fresh | Bull’s eye | 25 × 106 | 8 | 216 × 106 | 83% | 179 × 106 | 7.16 |
6 | Horse 3 | P4 | Frozen | Bull’s eye | 20 × 106 | 10 | 170 × 106 | 82% | 140 × 106 | 7 |
7 | Horse 4 | P5 | Frozen | Bull’s eye | 25 × 106 | 9 | 326 × 106 | 91% | 297 × 106 | 11.88 |
8 | Horse 5 | P4 | Frozen | Bull’s eye | 10 × 106 | 9 | 220 × 106 | 85% | 187 × 106 | 18.70 |
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Duysens, J.; Graide, H.; Niesten, A.; Mouithys-Mickalad, A.; Ceusters, J.; Serteyn, D. Optimization of the Amplification of Equine Muscle-Derived Mesenchymal Stromal Cells in a Hollow-Fiber Bioreactor. Methods Protoc. 2024, 7, 32. https://doi.org/10.3390/mps7020032
Duysens J, Graide H, Niesten A, Mouithys-Mickalad A, Ceusters J, Serteyn D. Optimization of the Amplification of Equine Muscle-Derived Mesenchymal Stromal Cells in a Hollow-Fiber Bioreactor. Methods and Protocols. 2024; 7(2):32. https://doi.org/10.3390/mps7020032
Chicago/Turabian StyleDuysens, Julien, Hélène Graide, Ariane Niesten, Ange Mouithys-Mickalad, Justine Ceusters, and Didier Serteyn. 2024. "Optimization of the Amplification of Equine Muscle-Derived Mesenchymal Stromal Cells in a Hollow-Fiber Bioreactor" Methods and Protocols 7, no. 2: 32. https://doi.org/10.3390/mps7020032