Mesodermal growth factor—A new type of cryoprotectant?

Weimar, Virginia; Haraguchi, Kenneth H.

doi:10.1016/0011-2240(79)90045-2

Cited by 4 publications

(2 citation statements)

References 3 publications

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“…The iris and lens were excised and 3 mm corneal buttons cut with a trephine. The buttons were soaked for 20 minutes in sterile organ culture medium prior to culturing (Weimar and Haraguchi, 1979b). The composition of the organ culture medium was Eagle's minimum essential medium (Earle's base) containing phenol red and supplemented with glutamine 2 mM; Eagle's nonessential amino acids, each 0.1 mM; penicillin 100 units/ml; streptomycin, 100 pg/ml; fungizone, 0.25 p.g/ml.…”

Section: Assay For Biological Activitymentioning

confidence: 99%

Synergistic growth stimulation of corneal fibroblasts by components of mesodermal growth factor from murine submaxillary glands

Haraguchi

Knox

Weimar

et al. 1982

Journal Cellular Physiology

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Recycled mesodermal growth factor (R-MGF) is a pool of proteins of 26,000 molecular weight obtained by recycling gel chromatography of male murine submaxillary gland extracts. R-MGF strikingly accelerates corneal stromal wound healing in vivo, fibroblast growth and migration in cultured corneal buttons and is showing here to stimulate stromal fibroblast growth and division in tissue culture. Chromatographic fractionation of R-MGF has yielded several components, none of which has a greater biological potency than the parent R-MGF. In contrast, two components, MGF-I and -II, when recombined synergistically stimulate fibroblast response in tissue culture and organ culture in excess of those obtained with the parent R-MGF. Three MGF components (I, III, and IV) have been purified and are inactive at 10-15 micrograms/ml in organ culture but potently stimulate fibroblast responses when combined in pairs containing 7.5 micrograms of each component. The striking synergism in organ culture suggests that the stimulation of wound healing by R-MGF in vivo may also reflect synergistic action of more than one R-MGF component. Procedures for isolating gram quantities of R-MGF and for the purification of different R-MGF components by ion exchange chromatography are detailed.

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Section: Assay For Biological Activitymentioning

confidence: 99%

Synergistic growth stimulation of corneal fibroblasts by components of mesodermal growth factor from murine submaxillary glands

Haraguchi

Knox

Weimar

et al. 1982

Journal Cellular Physiology

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“…Commonly immersion in a water bath of the vial containing the cornea and the freezing solution is used. Most authors use rapid thawing of the cornea at temperatures of at least +37¤°C [Weimar and Haraguchi, 1979].…”

Section: Introductionmentioning

confidence: 99%

Morphological Study of Cryopreserved Human Corneal Endothelium

Canals

Costa-Vila

Potau

et al. 1999

Cells Tissues Organs

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The aim of the present paper is to describe the morphological changes that occur in human corneal endothelium as an immediate consequence of corneal cryopreservation. Therefore, 16 human donor corneas were cryopreserved with an original procedure at a 1 °C/min cooling rate in a freezing solution cryoprotected with 7% dimethylsulphoxide until a final temperature of –100 °C was reached. After storage of the corneas in liquid nitrogen for periods ranging from 1 to 96 days (mean: 34.31 days), the corneas were thawed in a water bath at +37 °C. Eight additional control corneas were processed without cryopreservation. Morphological assessment of the endothelial layer was performed by scanning electron microscopy and trypan blue and alizarin red S vital staining. Results showed cryoinduced damage at variable degrees in all cryopreserved corneas. They were classified into three groups according to the intensity and extension of the cryoinduced damage: group I (n = 10): corneas with minor endothelial alterations consisting in the presence of microholes in the posterior cell membrane; group II (n = 1): corneas with generalized disruption of endothelial intercellular junctions and intact cell membranes; group III (n = 5): corneas with severe endothelial damage consisting of massive cell necrosis and complete alteration of the morphological pattern of the endothelium. All control corneas had intact endothelial layers. Cryoinduced damage cannot be completely avoided with the cryopreservation protocol tested. The high interindividual variability of the results observed is not related to the storage time of the cornea in liquid nitrogen.

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Canals

Garcı́a

Potau

et al. 2000

Cell and Tissue Banking

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The purpose of the present study was to set up and test a cryopreservation method for long-term storage of human corneas. Therefore the freezing solution was optimized in 264 rabbit corneas by testing the type of cryoprotectant, its concentration, addition and dilution pattern and exposure temperature. Then rabbit corneas were frozen in the optimum solution at different cooling rates and thawed in a water bath at different temperatures. Eight human corneas were cryopreserved with the method showing optimum results in rabbit corneas and four additional corneas were used as controls. Endothelial viability was assessed after each step by vital staining and scanning electron microscopy. Best results after exposure of rabbit corneas to the freezing solution were achieved when using a 10% cryoprotectant concentration, with direct addition/dilution and exposure at room temperature (3512 +/-300 viable cells mm(2) when using dimethylsulfoxide; 3403 +/- 245 viable cells mm(2) when using 1,2-propanediol). Cryopreserved rabbit corneas had the highest endothelial cell survival when frozen at 1 degrees C/min and thawed at 37 degrees C (2003 +/- 372 viable cells/mm(2) when using dimethylsulfoxide and 1357 +/- 667 viable cells/mm(2) when using 1,2-propanediol). Cryopreserved human corneas had 753 +/- 542 viable cells/mm(2) when using dimethylsulfoxide and 56 +/- 56 viable cells/mm(2) when using 1,2-propanediol. We can conclude that the method developed is easy to handle and shows optimum results in rabbit corneas, with an endothelial cell survival that is consistent with transplant acceptability criteria. The results obtained in human corneas are below prediction and are still unsatisfactory for successful use in eye banking.

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Mesodermal growth factor—A new type of cryoprotectant?

Cited by 4 publications

References 3 publications

Synergistic growth stimulation of corneal fibroblasts by components of mesodermal growth factor from murine submaxillary glands

Synergistic growth stimulation of corneal fibroblasts by components of mesodermal growth factor from murine submaxillary glands

Morphological Study of Cryopreserved Human Corneal Endothelium

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