Stimulation of corneal endothelial cell proliferation in vitro by fibroblast and epidermal growth factors

Gospodarowicz, Denis; Mescher, Anthony L.; Birdwell, Charles R.

doi:10.1016/0014-4835(77)90248-2

Cited by 326 publications

(148 citation statements)

References 19 publications

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“…Confluent cultures composed of flattened, closely apposed, and nonoverlapping cells were kept without further addition of FGF. Cultures of bovine corneal endothelial cells were established from steer eyes as described (16) and stock cultures maintained as in the case of vascular cells, except that 5% fetal calf serum was included in the growth medium.…”

Section: Methodsmentioning

confidence: 99%

Degradation of sulfated proteoglycans in the subendothelial extracellular matrix by human platelet heparitinase.

Yahalom¹,

Eldor²,

Fuks³

et al. 1984

J. Clin. Invest.

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As bstract. Cultured vascular and corneal endothelial cells produce an underlying extracellular matrix (ECM) which induces platelet adherence, aggregation, and release reaction. Incubation of a metabolically (35S)O--labeled ECM with platelet-rich plasma or washed platelets, but not with platelet-poor plasma, resulted in degradation of its heparan sulfate-containing proteoglycans into labeled fragments four to five times smaller than intact glycosaminoglycan side chains. These fragments were sensitive to deamination with nitrous acid and were not produced in the presence of heparin, indicating that heparan sulfate in the ECM is susceptible to cleavage by the platelet heparitinase. This degradation required adhesion of platelets to the ECM rather than aggregation since it was not inhibited by aspirin, which prevented platelet aggregation but not adherence. The enzyme was not released during aggregation of platelets on the ECM but was readily liberated upon their exposure to thrombin. This liberation was inhibited in the presence of prostacyclin (PGI2). Isolated high molecular weight proteoglycans first released from the ECM by incubation with platelet poor plasma served as a substrate for further degradation by the platelet heparitinase, suggesting a cascade mechanism for degradation of heparan sulfate in the ECM. Heparitinase, although to a lower level, was also active when washed platelets were added on top of a confluent endothelial cell monolayer covering the (35S)0=-labeled ECM. It is suggested that the platelet heparitinase may be involved in the impairment of the

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Section: Methodsmentioning

confidence: 99%

Degradation of sulfated proteoglycans in the subendothelial extracellular matrix by human platelet heparitinase.

Yahalom¹,

Eldor²,

Fuks³

et al. 1984

J. Clin. Invest.

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“…Cultures of bovine corneal endothelial (BCE) cells were established from steer eyes as previously described [20]. Stock cultures were maintained in DMEM (1 g/1 glucose) supplemented with 10% heat-inactivated newborn calf serum (NCS), 5% heat inactivated FCS, 50 U/ml penicillin, and 50 gtg/ml streptomycin at 37°C in a 10% CO2 humidified incubator.…”

Section: Cellsmentioning

confidence: 99%

The extracellular matrix produced by bovine corneal endothelial cells contains progelatinase A

Ménashi¹,

Vlodavsky²,

Ishai-Michaeli³

et al. 1995

FEBS Letters

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Progelatinase A is a matrix metalloproteinase involved in the turnover of extracellular matrix (ECM). We report that the ECM produced by bovine corneal endothelial (BCE) cells contains progelatinase A free of tissue inhibitor of metalloproteinase (TIMP2). The matrix-bound progelatinase A can be activated by APMA to generate a 62 kDa and a 45 kDa species with enzymatic activity and is inhibited by TIMP2. The bound progelatinase can be released after treatment of the ECM with gelatinase B. These studies suggest that the ECM can function as a reservoir for progelatinase A which may be readily available for cells in processes such as metastasis, angiogenesis, inflammation and wound heating.

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“…Chronic edema may result in vascularization of the stroma and a further loss of transparency. An (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.…”

mentioning

confidence: 99%

“…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.…”

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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Stimulation of corneal endothelial cell proliferation in vitro by fibroblast and epidermal growth factors

Cited by 326 publications

References 19 publications

Degradation of sulfated proteoglycans in the subendothelial extracellular matrix by human platelet heparitinase.

Degradation of sulfated proteoglycans in the subendothelial extracellular matrix by human platelet heparitinase.

The extracellular matrix produced by bovine corneal endothelial cells contains progelatinase A

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

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