Efficient Isolation and Functional Characterization of Niche Cells from Human Corneal Limbus
Abstract
:1. Introduction
2. Results
2.1. Localization of Limbal Niche Cells In Situ
2.2. Flow Sorting of Limbal Niche Cells
2.3. Characteristic Features of Sorted Cells
2.3.1. CD90+CD117− Cells (LMSC)
2.3.2. CD90−CD117+ Cells (LM)
2.3.3. CD90−CD117− Cells (LEPC)
2.3.4. The 3D Co-Cultures
3. Discussion
4. Materials and Methods
4.1. Cell Isolation
4.2. Fluorescence-Activated Cell Sorting (FACS)
4.3. Flow Cytometry
4.4. Growth Characteristics
4.4.1. Population Doubling Assay
4.4.2. Colony-Forming Unit Assay of LMSC and LM
4.5. Trilineage Differentiation
4.6. Melanin Production
4.7. Co-Culture Experiments
4.7.1. Colony-Forming Unit Assay of LEPC
4.7.2. The 3D Co-Cultures
4.8. Histology and Immunohistochemistry—Paraffin
4.9. Immunohistochemistry—Frozen and Immunocytochemistry
4.10. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Polisetti, N.; Sharaf, L.; Schlötzer-Schrehardt, U.; Schlunck, G.; Reinhard, T. Efficient Isolation and Functional Characterization of Niche Cells from Human Corneal Limbus. Int. J. Mol. Sci. 2022, 23, 2750. https://doi.org/10.3390/ijms23052750
Polisetti N, Sharaf L, Schlötzer-Schrehardt U, Schlunck G, Reinhard T. Efficient Isolation and Functional Characterization of Niche Cells from Human Corneal Limbus. International Journal of Molecular Sciences. 2022; 23(5):2750. https://doi.org/10.3390/ijms23052750
Chicago/Turabian StylePolisetti, Naresh, Lyne Sharaf, Ursula Schlötzer-Schrehardt, Günther Schlunck, and Thomas Reinhard. 2022. "Efficient Isolation and Functional Characterization of Niche Cells from Human Corneal Limbus" International Journal of Molecular Sciences 23, no. 5: 2750. https://doi.org/10.3390/ijms23052750