Modeling Traumatic Brain Injury in Human Cerebral Organoids
Abstract
:1. Introduction
2. Materials and Methods
2.1. Derivation and Characterization of iPSCs from Human Fibroblasts
2.2. Cerebral Organoid Generation
2.3. Animal Experiments
2.4. Controlled Cortical Impact Procedure in Live Mice
2.5. Phantom Brain Development
2.6. Mouse Skull Preparation for CCI
2.7. Controlled Cortical Impact Procedure in COs
2.8. Immunofluorescence
2.9. Statistical Analysis
3. Results
3.1. Generation and Characterization of the Phantom Brain
3.2. Generation and Characterization of Human iPSCs and COs
3.3. CCI Induces Astrogliosis and Reduces Neurons in COs
3.4. Elevated Neuronal Damage in COs after CCI
3.5. Cellular Apoptosis in COs after CCI
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
References
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Mix 1 | Mix 2 | Mix 3 | Mix 4 | Mix 5 | |
---|---|---|---|---|---|
Gelatin % | 0.6 | 0.6 | 0.8 | 1.5 | 1 |
Agarose % | 0.4 | 0.1 | 0.3 | 0.5 | 0.8 |
Neurons | For Each Organoid | All Together | |||||||
Mean | SD | CV | Mean | SD | CV | ||||
Org 1 | 315 | 324 | 343 | 327.33 | 14.295 | 4.367 | 333.87 | 13.98 | 4.18 |
Org 2 | 337 | 319 | 346 | 334 | 13.748 | 4.1161 | |||
Org 3 | 318 | 301 | 325 | 314.67 | 12.342 | 3.9224 | |||
Org 4 | 347 | 356 | 323 | 342 | 17.059 | 4.9879 | |||
Org 5 | 339 | 367 | 348 | 351.33 | 14.295 | 4.0686 | |||
Astrocytes | For Each Organoid | All Together | |||||||
Mean | SD | CV | Mean | SD | CV | ||||
Org 1 | 441 | 443 | 476 | 453.33 | 19.655 | 4.3357 | 507 | 52.4 | 10.34 |
Org 2 | 606 | 598 | 576 | 593.33 | 15.535 | 2.6182 | |||
Org 3 | 468 | 495 | 503 | 488.67 | 18.339 | 3.7529 | |||
Org 4 | 478 | 504 | 485 | 489 | 13.454 | 2.7513 | |||
Org 5 | 502 | 512 | 518 | 510.67 | 8.0829 | 1.5828 |
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Ramirez, S.; Mukherjee, A.; Sepulveda, S.; Becerra-Calixto, A.; Bravo-Vasquez, N.; Gherardelli, C.; Chavez, M.; Soto, C. Modeling Traumatic Brain Injury in Human Cerebral Organoids. Cells 2021, 10, 2683. https://doi.org/10.3390/cells10102683
Ramirez S, Mukherjee A, Sepulveda S, Becerra-Calixto A, Bravo-Vasquez N, Gherardelli C, Chavez M, Soto C. Modeling Traumatic Brain Injury in Human Cerebral Organoids. Cells. 2021; 10(10):2683. https://doi.org/10.3390/cells10102683
Chicago/Turabian StyleRamirez, Santiago, Abhisek Mukherjee, Sofia Sepulveda, Andrea Becerra-Calixto, Nicolas Bravo-Vasquez, Camila Gherardelli, Melissa Chavez, and Claudio Soto. 2021. "Modeling Traumatic Brain Injury in Human Cerebral Organoids" Cells 10, no. 10: 2683. https://doi.org/10.3390/cells10102683