December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Immunohistological and Biochemical Characterization of Cultured MMP-Deficient Mouse Corneal Epithelial Cells
CC Chen; JH Chang; J Savage; MA Shatos; E Gabison; D Kure; DT Azar
Author Affiliations & Notes
  • CC Chen
    Ophthalmology Massachusetts Eye and Ear Infirmary Schepens Eye Research Institute Harvard Medical School Boston MA
  • JH Chang
    Ophthalmology Massachusetts Eye and Ear Infirmary Schepens Eye Research Institute Harvard Medical School Boston MA
  • J Savage
    Ophthalmology Massachusetts Eye and Ear Infirmary Schepens Eye Research Institute Harvard Medical School Boston MA
  • MA Shatos
    Ophthalmology Massachusetts Eye and Ear Infirmary Schepens Eye Research Institute Harvard Medical School Boston MA
  • E Gabison
    Ophthalmology Massachusetts Eye and Ear Infirmary Schepens Eye Research Institute Harvard Medical School Boston MA
  • D Kure
    Ophthalmology Massachusetts Eye and Ear Infirmary Schepens Eye Research Institute Harvard Medical School Boston MA
  • DT Azar
    Ophthalmology Massachusetts Eye and Ear Infirmary Schepens Eye Research Institute Harvard Medical School Boston MA
  • Footnotes
    Commercial Relationships   C.C. Chen, None; J.H. Chang, None; J. Savage, None; M.A. Shatos, None; E. Gabison, None; D. Kure, None; D.T. Azar, None. Grant Identification: NIH EY 10101
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 1759. doi:
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      CC Chen, JH Chang, J Savage, MA Shatos, E Gabison, D Kure, DT Azar; Immunohistological and Biochemical Characterization of Cultured MMP-Deficient Mouse Corneal Epithelial Cells . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1759.

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Abstract

Abstract: : Purpose: To generate and characterize immortalized corneal epithelial and keratocyte cell lines from matrix metalloproteinase (MMP)-deficient mice. Methods: Mouse corneas were obtained immediately after enucleation and the epithelial and stromal layers were separated under a dissecting microscope. They were cultured to generate primary and SV40-immortalized corneal cells derived from MMP-2, -3, -7, -12 deficient (MMP-/-) and wild type C57BL/6 mice. We determined the immunoreactivity of the cell lines with the following antibodies: AE1/AE3 (for most acidic keratins and known basic keratins), J7 (for 55-kD cytokeratin), vimentin, type VIII collagen (for endothelial cells), α-smooth muscle actin (α-SMA; for myofibroblasts). The cytoskeletal organization and co-localization were evaluated using epifluorescent and confocal microscopy. In addition, cellular extracts of MMP -/- and wild type mice were used to compare their inhibitory effect on vascular endothelial cell proliferation. ANOVA and student's t test were used for statistical analysis. Results: Of the epithelial and keratocyte mouse cell lines derived from MMP-/- mice (designated as MSM-E, MSM-K, respectively) and wild type mice (designated as MSW-TE, MSW-TK, respectively), the MSM-2E, MSM-3E, MSM-7E and MSW-TE exhibited uniformly polygonal-shaped cells characteristic of stratified squamous epithelium. All MSM-E and MSW-TE cell lines expressed AE1/AE3, J7 and vimentin, but not type VIII collagen and α-SMA. In contrast, MSM-K (MSM-2K, MSM-3K, MSM-7K, MSM-12K) and MSW- TK cell lines were elongated, spindle shaped with scattered whorl formation and expressed vimentin and α-SMA. Cellular extracts of MSM-7E showed significant inhibition of calf pulmonary arterial endothelial cell proliferation in vitro (P<0.05). No significant differences in proliferation were observed between MSM-7E and MSW-TE cell lines. Conclusion: The MSM-E and MSM-K mouse cell lines exhibited in vitro characteristics of corneal epithelial cells and keratocytes, respectively. MSM-E and MSM-K cell lines may be useful in studying corneal wound healing, angiogenesis and other corneal pathologic and biologic processes.

Keywords: 370 cornea: basic science • 403 extracellular matrix • 434 immunohistochemistry 
© 2002, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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