Osmotic properties of the rabbit corneal endothelium and their relevance to cryopreservation |
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Authors: | D E Pegg C J Hunt L P Fong |
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Institution: | (1) MRC Medical Cryobiology Group, University Department of Surgery, Douglas House, Trumpington Road, CB2 2AH Cambridge, UK;(2) Present address: Hilles Immunology Laboratory, Harvard Medical School, 243 Charles Street, 021114 Boston, MA |
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Abstract: | The process of cryopreservation subjects cells to gross changes in the composition of the solution that surrounds them, changes
that cause the cells first to shrink and then to swell by an osmotic mechanism. Empirical methods have been developed that
permit many cells to survive freezing and thawing, but the cornea, which is crucially dependent upon the function of its endothelial
monolayer, has proved quite refractory. In this paper we explore the osmotic response of the corneal endothelium of the rabbit
to solutions ranging in osmolality from 0.25 to 8.6 × isotonic. Boyle van't Hoff behavior was observed between 0.43 and 8.6
× isotonic, and there was an apparent nonosmotic volume of 33.6%. However, ultrastructural damage was observed at the limits
of this range, and it appeared that the tolerated range was 0.64–4.4 × isotonic. We show the extent to which dimethyl sulfoxide
(Me2SO) would be expected to moderate changes in volume during freezing and suggest that its initial concentration should be at
least 2M to prevent excessive shrinkage. We also show that cell swelling during removal of Me2SO is especially likely to be hazardous. |
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Keywords: | Corneal endothelium Boyle van't Hoff response cryopreservation osmotic properties of corneal endothelium |
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