Preclinical Efficacy of Peripheral Nerve Regeneration by Schwann Cell-like Cells Differentiated from Human Tonsil-Derived Mesenchymal Stem Cells in C22 Mice
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cultivation of TMSCs and NRPCs
2.2. Animal Experiments
2.2.1. C22 Mice
2.2.2. Transplantation
2.2.3. Gait Test
2.2.4. Rotarod Test
2.2.5. Nerve Conduction Study
2.3. Transmission Electron Microscopy
2.4. Immunohistochemistry
2.5. Fluorescence In Situ Hybridization
2.6. Real-Time Quantitative Polymerase Chain Reaction
2.7. Statistical Analysis
3. Results
3.1. NRPCs Improved Behavioral Test Results in a Single-Dose Transplantation Experiment
3.2. Restoration of Sciatic Nerve Structure in a Single-Dose Transplantation Experiment
3.3. Regeneration of the Sciatic Nerve and Gastrocnemius and the Related Pathway in a Single-Dose Transplantation Experiment
3.4. NRPC Improved Behavioral Test Results in the Repeated-Dose Transplantation Experiment
3.5. Restoration of the Sciatic Nerve Structure in the Repeated-Dose Transplantation Experiment
3.6. Restoration of the Sciatic Nerve and Gastrocnemius in the Repeated-Dose Transplantation Experiment
3.7. Downregulation of PMP22 and the Presence of NRPCs 12 Weeks after Repeated-Dose Transplantation in the Sciatic Nerve
3.8. Expression of Peripheral Nerve Regeneration-Related microRNA in NRPCs
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Characteristics | Value/Specification |
---|---|
Viability | >90% |
Morphology | Specific form for neural precursor cell of the shape of a bipolar or tripolar |
Positive markers | CD73, CD90, CD105 |
Negative markers | CD14, CD34, CD45 |
Infectious agent test | SCB (p4), WCB (p9), F/P (p14) |
Other quality tests | HIV RNA, HCV RNA, HBV DNA, CMV RNA, anti-HIV, anti-HCV, anti-HBsAg, anti-CMV IgM/IgG, anti-HTLV-1/2, Syphilis |
Cryoprotectant | Cryostor CS10 |
Experiment | Group | Number of Cells for Transplantation | Mouse | Number of Animals (Male) | Route of Administration | Dosing Frequency |
---|---|---|---|---|---|---|
Single administration | NRPC-low | 2.5 × 104 cells/ 100 μL/site × 2 sites | C22 mice | 11 |
| Single |
NRPC-med | 2.5 × 105 cells/ 100 μL/site × 2 sites | 11 | ||||
NRPC-high | 5.0 × 105 cells/ 100 μL/site × 2 sites | 12 | ||||
Sham | CS10/100 μL/ site × 2 sites | 11 | ||||
W/T | - | Wild-type mice | 12 | |||
Repeated administration | NRPC-low | 2.5 × 104 cells/ 100 μL/site × 2 sites | C22 mice | 11 |
| Two repetitions 4 weeks apart |
NRPC-med | 2.5 × 105 cells/ 100 μL/site × 2 sites | 11 | ||||
NRPC-high | 5.0 × 105 cells/ 100 μL/site × 2 sites | 11 | ||||
Sham | CS10/100 μL/ site × 2 sites | 11 | ||||
W/T | - | Wild-type mice | 13 |
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Nam, Y.H.; Park, S.; Yum, Y.; Jeong, S.; Park, H.E.; Kim, H.J.; Lim, J.; Choi, B.-O.; Jung, S.-C. Preclinical Efficacy of Peripheral Nerve Regeneration by Schwann Cell-like Cells Differentiated from Human Tonsil-Derived Mesenchymal Stem Cells in C22 Mice. Biomedicines 2023, 11, 3334. https://doi.org/10.3390/biomedicines11123334
Nam YH, Park S, Yum Y, Jeong S, Park HE, Kim HJ, Lim J, Choi B-O, Jung S-C. Preclinical Efficacy of Peripheral Nerve Regeneration by Schwann Cell-like Cells Differentiated from Human Tonsil-Derived Mesenchymal Stem Cells in C22 Mice. Biomedicines. 2023; 11(12):3334. https://doi.org/10.3390/biomedicines11123334
Chicago/Turabian StyleNam, Yu Hwa, Saeyoung Park, Yoonji Yum, Soyeon Jeong, Hyo Eun Park, Ho Jin Kim, Jaeseung Lim, Byung-Ok Choi, and Sung-Chul Jung. 2023. "Preclinical Efficacy of Peripheral Nerve Regeneration by Schwann Cell-like Cells Differentiated from Human Tonsil-Derived Mesenchymal Stem Cells in C22 Mice" Biomedicines 11, no. 12: 3334. https://doi.org/10.3390/biomedicines11123334