Endothelial wound repair in primate cornea

Horn, Diane L. Van; Hyndiuk, Robert A.

doi:10.1016/0014-4835(75)90076-7

Cited by 168 publications

(70 citation statements)

References 15 publications

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“…One possible explanation may be that the cells readily obtained a differentiated phenotype when they were maintained in DMEM/10% FBS medium. It is, however, intriguing to note that despite the apparent diverse proliferative potential of the CE in different mammals [1,[53][54][55], the CE spheres having been isolated from human, rabbits, and cows appear to share a highly conserved phenotype.…”

Section: Discussionmentioning

confidence: 99%

Bovine Posterior Limbus: An Evaluation of an Alternative Source for Corneal Endothelial and Trabecular Meshwork Stem/Progenitor Cells

Grierson

Sheridan

et al. 2015

Stem Cells and Development

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A growing body of evidence has revealed that stem-like cells in the posterior limbus of the eye between the corneal endothelium (CE) and trabecular meshwork (TM) may be able to rejuvenate these tissues in disease. However, these cells have not been clearly defined and we have named them PET cells (progenitor cells of the endothelium and trabeculum). A good and inexpensive animal model for PET cells is lacking, so we investigated bovine eyes as an effective large tissue source. We showed the presence of stem/progenitor cells in the bovine CE, transition zone, and TM in situ. Floating spheres cultured from the CE and TM showed similar stem cell marker expression patterns. Both the CE and TM spheres were bipotent and highly proliferative, but with limited secondary sphere-forming capability. They were highly prone to differentiate back into the cell type of their tissue of origin. It is speculated that the PET cells become more tissue-specific as they migrate away from their niche. Here, we showed that PET cells are present in the posterior limbus of bovine eyes and that they can be successfully cultured and expanded. PET cells represent an attractive target for developing new treatments to regenerate both the CE and TM, thereby reducing the requirement for donor tissue for corneal transplant and invasive treatments for glaucomatous patients.

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Section: Discussionmentioning

confidence: 99%

Bovine Posterior Limbus: An Evaluation of an Alternative Source for Corneal Endothelial and Trabecular Meshwork Stem/Progenitor Cells

Grierson

Sheridan

et al. 2015

Stem Cells and Development

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“…Only rarely has proliferation been observed in the adult human cornea in vivo ( 1). In many mammalian species, a diminished central corneal endothelial density is observed in the adult animal owing to limited regeneration and enlargement of the eye (2,3). The limited regeneration is insufficient to compensate for irreversible damage which may occur after inflammation or surgical trauma.…”

Section: Introductionmentioning

confidence: 99%

Expression of SV40 virus large T antigen by recombinant adenoviruses activates proliferation of corneal endothelium in vitro.

Feldman¹,

Gjerset²,

Gately³

et al. 1993

J. Clin. Invest.

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Infection with the Ad5-SVR4 virus was used to introduce the large T antigen encoding region of the SV40 virus into bovine and human corneal endothelial cells. Expression of large T antigen occurred in 40% of bovine corneal endothelial cells after a 24-h incubation time versus 12% after 8 h of incubation. By 48 h after infection, almost all (92.8%) bovine corneal endothelial cells expressed large T antigen. Bovine and human corneal endothelial cells which expressed large T antigen proliferated and the characteristic morphologic features of corneal endothelium were maintained. This method may enable growth of enough corneal endothelium to perform studies to elucidate the biochemical mechanisms involved in regulating endothelial cell function. (J. Clin. Invest. 1993. 91:1713-1720

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“…The relative dehydration of the corneal tissue is determined primarily by the active transport pump of the corneal endothelium. The ability of most corneal endothelia to repair themselves in vivo is severely limited by their lack of proliferative capacity (1,2). Although small endothelial wounds can be repaired through the process of endomitosis and cell enlargement (3), extensive wounds may result in swelling of the corneal stroma and loss of transparency.…”

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confidence: 99%

“…The cell suspension was then centrifuged, the supernatant was aspirated, and the cell pellet was resuspended in medium/0.1% calf serum at a final concentration of 105 cells per 70 ,l; 120 ,l was added to each cornea. The corneas were incubated in a water-jacketed humidified CO2 incubator for [2][3][4][5][6][7][8][9][10][11][12] hr. Evaluation of the plating efficiency and of the cellular morphology was done by using alizarin red staining of the cornea (6)(7)(8)(9)(10)(11).…”

mentioning

confidence: 99%

“…DISCUSSION Rabbits were used in these studies, for reasons of economy and availability. Because it'is known that the corneal endothelium of the rabbit, unlike that of other species such as felines or primates, retains its ability to divide and to regenerate after trauma in vivo (2,(15)(16)(17), it is imperative to establish that the clarity of the graft was not due to a recolonization of the graft by the recipient rabbit corneal endothelial cells. That this was not the case can be concluded from four lines of evidence: (i) After keratoplasty, the graft containing bovine corneal endothelial cells remained clear from the outset.…”

mentioning

confidence: 99%

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Transplantation of cultured bovine corneal endothelial cells to rabbit cornea: clinical implications for human studies.

Gospodarowicz¹,

Greenburg²,

Alvarado³

1979

Proc. Natl. Acad. Sci. U.S.A.

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Rabbit corneas denuded of their endothelium were coated with bovine corneal endothelial cells (from steers) previously maintained in tissue culture for short (20 generations) or prolonged (200 generations) periods. When grafted back into female rabbits, the corneal buttons remained clear and showed no edema. In contrast, denuded corneas coated with bovine keratocytes and grafted into rabbits became opaque and edematous within 7 da s and remained so thereafter. Bovine corneal endothelial cells of the grafted corneas, which had remained clear for over 100 days, proliferated actively when put back into tissue culture. The corneal endothelial cells of the graft had a chromosome number of 2n = 60. The sex chromosomes were characteristic of the male (XY). The chromosome number of the endothelium of the recipient rabbit was 2n = 44 with sex chromosomes characteristic of the female (XX). Results of the karyotype analysis show that there was no invasion of the corneal button by the recipient endothelium and, conversely, no invasion of the recipient endothelium by the endothelium on the corneal button. These results demonstrate that cultured corneal endothelial cells remain functional in vitro and can replace a damaged or nonfunctional endothelium in vivo. The cornea owes its transparency to its uniform structure, avascularity, and deturgescence. The relative dehydration of the corneal tissue is determined primarily by the active transport pump of the corneal endothelium. The ability of most corneal endothelia to repair themselves in vivo is severely limited by their lack of proliferative capacity (1, 2). Although small endothelial wounds can be repaired through the process of endomitosis and cell enlargement (3) (5). The cells were maintained on gelatinized dishes in Dulbecco's modified Eagle's medium supplemented with 10% calf serum, 5% dextran, 50 ,ug of gentamycin per ml, and 2.5 .g of Fungizone per ml. Fibroblast growth factor (100 ng/ml) was added every other day (5, 6). The cultures were passaged every week with a split ratio of 1:64. The morphological and biochemical characteristics of these cultures have been described (5-10). Cultures of bovine corneal keratocytes were established as described (7).Coating of Denuded Descemet's Membrane of Rabbit Cornea with Bovine Corneal Endothelial Cells or Keratocytes. Female Dutch rabbits were used throughout this study. The procedure is summarized in Fig. 1. After being anesthetized, the animal was sacrificed and the eye was enucleated. The cornea and a rim of sclera were then excised. The cornea was placed in a petri dish so that the endothelial layer faced upward. A cotton swab was used to sweep the surface of the cornea (6). [Control studies with an alizarin red staining technique have shown that the entire endothelial cell layer is removed and that no damage is done to Descemet's membrane (6,9).] A corneal button was cut with an 1 -mm trephine and placed in a Teflon container fashioned to fit the contour of the cornea. Concurrently with the dissection and pr...

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Endothelial wound repair in primate cornea

Cited by 168 publications

References 15 publications

Bovine Posterior Limbus: An Evaluation of an Alternative Source for Corneal Endothelial and Trabecular Meshwork Stem/Progenitor Cells

Bovine Posterior Limbus: An Evaluation of an Alternative Source for Corneal Endothelial and Trabecular Meshwork Stem/Progenitor Cells

Expression of SV40 virus large T antigen by recombinant adenoviruses activates proliferation of corneal endothelium in vitro.

Transplantation of cultured bovine corneal endothelial cells to rabbit cornea: clinical implications for human studies.

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