Investigating the Role of TGF-β Signaling Pathways in Human Corneal Endothelial Cell Primary Culture
Abstract
:1. Introduction
2. Materials and Methods
2.1. Ethical Statement
2.2. Crucial Products
2.3. Primary Culture of hCECs
2.4. Immunofluorescence (IF)
2.5. Methods of Quantification
2.6. Statistics
3. Results
3.1. The Three TGF-β Isforms Induced EndMT on Confluent hCECs in Absence of FBS
3.2. Selection of NCAM as Biomarker to Assess EndMT
3.3. TGF-β1 and 3 Induced EndMT and Lower ECD on Cultured hCECs in Presence of FBS
3.4. The Assessment of Inhibitors of TGF-β Signaling Pathways Revealed Positive Effects in Inhibiting P38-MAPK, ROCK, and TGF-β Receptor
4. Discussion
4.1. Effects of TGF-β 1, 2, and 3 on In Vitro hCECs
4.2. Proposal of a Screening System for Molecules to Evaluate Their Effects on hCECs In Vitro
4.3. Effects of Inhibiting Various TGF-β Signaling Pathways on Cultured hCECs
4.4. Limitations and Perspectives
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Inhibitor | Catalog Number | Provider | Final Concentration | Signaling Pathway Inhibition | Reference |
---|---|---|---|---|---|
SIS3 HCL | S7959 | SelleckChem | 3 µM | SMAD 2/3 | [30] |
Asiaticoside | HY N0439 | MedChemExpress | 100 µg/mL | SMAD 2/3 | [31] |
Y27632 | S1049 | SelleckChem | 10 µM | Rho/ROCK | [32] |
Ripasudil | HY 15685 | MedChemExpress | 5 µM | Rho/ROCK | [33] |
Tectochrysin | HY 14592 | MedChemExpress | 10 µg/mL | NF-κB | [34] |
QNZ | S4902 | SelleckChem | 0.1 µM | NF-κB | [35] |
MK-2206 | S107811 | SelleckChem | 10 µM | PI3K/AKT/mTOR | [36] |
LY294002 | L9908 | Sigma-Aldrich | 20 µM | PI3K/AKT/mTOR | [37] |
SB203580 | S1076 | SelleckChem | 10 µM | P38 MAPK | [38] |
SB202190 | SYN-1073 | SYNkinase | 10 µM | P38 MAPK | [39] |
U0126 | 6620056 | CalbioChem | 10 µM | Ras/Raf/MEK/ERK | [40] |
AZD0364 | S8708 | SelleckChem | 2 µM | Ras/Raf/MEK/ERK | [41] |
SB431542 | S1067 | SelleckChem | 10 µM | TGFB receptor | [28] |
LY2109761 | SML 2051 | SelleckChem | 1 µM | TGFB receptor | [27] |
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Share and Cite
Aouimeur, I.; Sagnial, T.; Coulomb, L.; Maurin, C.; Thomas, J.; Forestier, P.; Ninotta, S.; Perrache, C.; Forest, F.; Gain, P.; et al. Investigating the Role of TGF-β Signaling Pathways in Human Corneal Endothelial Cell Primary Culture. Cells 2023, 12, 1624. https://doi.org/10.3390/cells12121624
Aouimeur I, Sagnial T, Coulomb L, Maurin C, Thomas J, Forestier P, Ninotta S, Perrache C, Forest F, Gain P, et al. Investigating the Role of TGF-β Signaling Pathways in Human Corneal Endothelial Cell Primary Culture. Cells. 2023; 12(12):1624. https://doi.org/10.3390/cells12121624
Chicago/Turabian StyleAouimeur, Inès, Tomy Sagnial, Louise Coulomb, Corantin Maurin, Justin Thomas, Pierre Forestier, Sandrine Ninotta, Chantal Perrache, Fabien Forest, Philippe Gain, and et al. 2023. "Investigating the Role of TGF-β Signaling Pathways in Human Corneal Endothelial Cell Primary Culture" Cells 12, no. 12: 1624. https://doi.org/10.3390/cells12121624