April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Inmunologic Response of the Rabbit Cornea to Xenogenic and Allogeneic Bone Marrow Derived Mesenchymal Stem Cells (MSC)
C. Nien Shy; T. Yamagami; K. Flynn; D. Carpenter; H. Levis; J. Simpson; J. V. Jester
Author Affiliations & Notes
  • C. Nien Shy
    The Gavin S. Herbert Eye Institute, University of California, Irvine, Orange, California
  • T. Yamagami
    The Gavin S. Herbert Eye Institute, University of California, Irvine, Orange, California
  • K. Flynn
    The Gavin S. Herbert Eye Institute, University of California, Irvine, Orange, California
  • D. Carpenter
    The Gavin S. Herbert Eye Institute, University of California, Irvine, Orange, California
  • H. Levis
    The Gavin S. Herbert Eye Institute, University of California, Irvine, Orange, California
  • J. Simpson
    The Gavin S. Herbert Eye Institute, University of California, Irvine, Orange, California
  • J. V. Jester
    The Gavin S. Herbert Eye Institute, University of California, Irvine, Orange, California
  • Footnotes
    Commercial Relationships  C. Nien Shy, None; T. Yamagami, None; K. Flynn, None; D. Carpenter, None; H. Levis, None; J. Simpson, None; J.V. Jester, None.
  • Footnotes
    Support  EY016663, Research to Prevent Blindness, Cystinosis Research Foundation
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 1772. doi:
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      C. Nien Shy, T. Yamagami, K. Flynn, D. Carpenter, H. Levis, J. Simpson, J. V. Jester; Inmunologic Response of the Rabbit Cornea to Xenogenic and Allogeneic Bone Marrow Derived Mesenchymal Stem Cells (MSC). Invest. Ophthalmol. Vis. Sci. 2009;50(13):1772.

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Abstract

Purpose: : Adult MSC are multipotent progenitor cells capable to differentiating into different cell types, including corneal stromal cells. The purpose of this study was to characterize the basic immunological responses of the rabbit cornea to xenogenic human and mouse MSC (h-MSC and m-MSC respectively) and compare to allogeneic rabbit MSC (r-MSC) and corneal fibroblasts (RCF) to provide a basis for developing therapeutic strategies for treating corneal diseases.

Methods: : GFP expressing h-MSC and m-MSC were obtained from Tulane University. R-MSC cultures were established using plastic adherence of bone marrow cells obtained from 3 month old New Zealand albino rabbit and maintained in -MEM. The differentiation potential of r-MSC was confirmed by culturing in differentiation media specific for osteogenesis and adipogenesis followed by Alizarin Red and Oil-Red-O staining respectively. RCF were established from collagenase digested corneas and maintained in 10% serum supplemented DMEM. Both r-MSC and RCF were later transfected with a feline leukemia virus vector expressing GFP driven by the mouse stem cell promoter. Up to 50,000 cells in 25µl of PBS were injected into the corneal stroma of anesthetized rabbits using a 30G needle. Animals were then sacrifice 7 days after injection and corneas collected for inmunostaining with CD45, CD4, CD8 and macrophage (RAM11). In addition, eyes were scanned using in vivo and ex vivo confocal microscopy to asses corneal haze and localization of GFP expressing cells.

Results: : Injected h-MSC, m-MSC and r-MSC maintained a rounded morphology in situ and showed extensive light scattering. RCF showed a spindle shaped morphology but also exhibited extensive light scattering. Immunostaining detected marked infiltration of leukocytes staining for CD4, CD8 and RAM11 in h-MSC and m-MSC injected corneas. RCF injected corneas showed substantially reduced leukocyte infiltration while r-MSC showed only presence of macrophages.

Conclusions: : Our results suggest that xenogenic MSC induced immune responses including lymphocytic infiltration of CD4 and CD8 T-cells with macrophage recruitment. Allogeneic RCF induced milder immune response while allogeneic MSC showed only recruitment of macrophages. We concluded that future studies should use allogeneic MSC to evaluate corneal keratocyte differentiation potential of MSC.

Keywords: cornea: stroma and keratocytes • transplantation • immunohistochemistry 
© 2009, 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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