Differentiation-related expression of a major 64K corneal keratin in vivo and in culture suggests limbal location of corneal epithelial stem cells

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Differentiation-related expression of a major 64K corneal keratin in vivo and in culture suggests limbal location of corneal epithelial stem cells

A Schermer, S Galvin and TT Sun

In this paper we present keratin expression data that lend strong support to a model of corneal epithelial maturation in which the stem cells are located in the limbus, the transitional zone between cornea and conjunctiva. Using a new monoclonal antibody, AE5, which is highly specific for a 64,000-mol-wt corneal keratin, designated RK3, we demonstrate that this keratin is localized in all cell layers of rabbit corneal epithelium, but only in the suprabasal layers of the limbal epithelium. Analysis of cultured corneal keratinocytes showed that they express sequentially three major keratin pairs. Early cultures consisting of a monolayer of "basal" cells express mainly the 50/58K keratins, exponentially growing cells synthesize additional 48/56K keratins, and postconfluent, heavily stratified cultures begin to express the 55/64K corneal keratins. Cell separation experiments showed that basal cells isolated from postconfluent cultures contain predominantly the 50/58K pair, whereas suprabasal cells contain additional 55/64K and 48/56K pairs. Basal cells of the older, postconfluent cultures, however, can become AE5 positive, indicating that suprabasal location is not a prerequisite for the expression of the 64K keratin. Taken together, these results suggest that the acidic 55K and basic 64K keratins represent markers for an advanced stage of corneal epithelial differentiation. The fact that epithelial basal cells of central cornea but not those of the limbus possess the 64K keratin therefore indicates that corneal basal cells are in a more differentiated state than limbal basal cells. These findings, coupled with the known centripetal migration of corneal epithelial cells, strongly suggest that corneal epithelial stem cells are located in the limbus, and that corneal basal cells correspond to "transient amplifying cells" in the scheme of "stem cells----transient amplifying cells----terminally differentiated cells."
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Hong, K. U., Reynolds, S. D., Giangreco, A., Hurley, C. M., Stripp, B. R. (2001). Clara Cell Secretory Protein-Expressing Cells of the Airway Neuroepithelial Body Microenvironment Include a Label-Retaining Subset and Are Critical for Epithelial Renewal after Progenitor Cell Depletion. Am. J. Respir. Cell Mol. Bio. 24: 671-681 [Abstract] [Full Text]
Anderson, D. F, Ellies, P., Pires, R. T F, Tseng, S. C G (2001). Amniotic membrane transplantation for partial limbal stem cell deficiency. Br J Ophthalmol 85: 567-575 [Abstract] [Full Text]
Anderson, S. C., SundarRaj, N. (2001). Regulation of a Rho-Associated Kinase Expression during the Corneal Epithelial Cell Cycle. IOVS 42: 933-940 [Abstract] [Full Text]
Pellegrini, G., Dellambra, E., Golisano, O., Martinelli, E., Fantozzi, I., Bondanza, S., Ponzin, D., McKeon, F., De Luca, M. (2001). p63 identifies keratinocyte stem cells. Proc. Natl. Acad. Sci. USA 98: 3156-3161 [Abstract] [Full Text]
Nakamura, T., Nishida, K., Dota, A., Matsuki, M., Yamanishi, K., Kinoshita, S. (2001). Elevated Expression of Transglutaminase 1 and Keratinization-Related Proteins in Conjunctiva in Severe Ocular Surface Disease. IOVS 42: 549-556 [Abstract] [Full Text]
ELLIES, P., ANDERSON, D. F, TOUHAMI, A., TSENG, S. C G (2001). Limbal stem cell deficiency arising from systemic chemotherapy. Br J Ophthalmol 85: 371c-371 [Full Text]
Koizumi, N., Inatomi, T., Suzuki, T., Sotozono, C., Kinoshita, S. (2001). Cultivated Corneal Epithelial Transplantation for Ocular Surface Reconstruction in Acute Phase of Stevens-Johnson Syndrome. Arch Ophthalmol 119: 298-300 [Full Text]
Taliana, L., Evans, M. D. M., Dimitrijevich, S. D., Steele, J. G. (2001). The Influence of Stromal Contraction in a Wound Model System on Corneal Epithelial Stratification. IOVS 42: 81-89 [Abstract] [Full Text]
Yoshida, K., Hu, Y., Karin, M. (2000). I{kappa}B Kinase {alpha} Is Essential for Development of the Mammalian Cornea and Conjunctiva. IOVS 41: 3665-3669 [Abstract] [Full Text]
Daniels, J. T., Khaw, P. T. (2000). Temporal Stimulation of Corneal Fibroblast Wound Healing Activity by Differentiating Epithelium In Vitro. IOVS 41: 3754-3762 [Abstract] [Full Text]
Ikeda, T., Ishii, H., Higuchi, T., Sato, K., Hayashi, Y., Ikeda, K., Hirabayashi, Y. (2000). Localization of Thrombomodulin in the Anterior Segment of the Human Eye. IOVS 41: 3383-3390 [Abstract] [Full Text]
Wang, I-J., Hu, F.-R., Chen, P.-J., Lin, C.-T. (2000). Mechanism of Abnormal Elastin Gene Expression in the Pinguecular Part of Pterygia. Am. J. Pathol. 157: 1269-1276 [Abstract] [Full Text]
Koizumi, N., Fullwood, N. J., Bairaktaris, G., Inatomi, T., Kinoshita, S., Quantock, A. J. (2000). Cultivation of Corneal Epithelial Cells on Intact and Denuded Human Amniotic Membrane. IOVS 41: 2506-2513 [Abstract] [Full Text]
Philipp, W., Speicher, L., Humpel, C. (2000). Expression of Vascular Endothelial Growth Factor and Its Receptors in Inflamed and Vascularized Human Corneas. IOVS 41: 2514-2522 [Abstract] [Full Text]