Organ culture: the method of choice for preservation of human donor corneas

Pels, Liesbeth

doi:10.1136/bjo.81.7.523

Cited by 89 publications

(72 citation statements)

References 28 publications

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“…Glycerol is a dehydrating agent with antimicrobial and antiprotease properties. However, glycerol will not preserve endothelial viability, a condition needed for optical PKP; cryopreservation 12,13 and 34 1C organ culture 14 will allow long-term storage of viable corneal tissue. Cryopreservation and organ culture, because of their technical complexity and cost, have limited application in the areas, which lack fresh donor corneas.…”

Section: Resultsmentioning

confidence: 99%

Eye preservation tectonic graft using glycerol-preserved donor cornea

Hc¹,

Sj²,

Chao³

2012

Eye

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Aims To report the surgical outcome of tectonic graft using glycerol-preserved donor corneas to treat perforated keratitis. Methods The medical records were reviewed of all patients treated for perforated keratitis using glycerol-preserved corneas at a single institution between 1 July 2004 and 31 June 2010. The clinical features, precipitating factors, adjuvant therapies, and therapeutic outcomes were analyzed. Success was defined as re-epithelialization of the ocular surface without evisceration. Results Fourteen eyes from 14 patients (6 male and 8 female) were included. Age ranged from 58 to 84 years (average, 70.71±8.52 years) and the follow-up time ranged from 7 to 56 months (mean, 25.35 ± 16.84 months). The culture results showed five bacterial infections, five cases of fungal keratitis, and one mixed infection; the culture results were negative for three patients. Satisfactory anatomical integrity was obtained in eight grafts (57.14%) that healed with neovascularization. Six grafts (48.85%) showed delayed re-epithelialization and were repaired with conjunctival flaps to maintain ocular surface integrity. Three patients developed secondary glaucoma and received trans-scleral cyclophotocoagulation. Thirteen patients had satisfactory anatomical integrity without evisceration or exenteration, while one patient received evisceration at 39-month follow-up because of intractable glaucoma. Conclusions Glycerol-preserved donor corneas combined with anterior vitrectomy with or without conjunctival flaps may be effective substitutes for evisceration surgery in patients with perforated keratitis.

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Section: Resultsmentioning

confidence: 99%

Eye preservation tectonic graft using glycerol-preserved donor cornea

Hc¹,

Sj²,

Chao³

2012

Eye

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“…They were not suitable for transplantation owing to inconclusive donor serology despite normal endothelial characteristics, in particular central ECD >2,000 cells per mm 2 , the conventional cutoff value for corneal graft qualification. They were stored in organ culture (OC), the prevalent method used in European eye banks, which maintains cell metabolic activity for at least 5 weeks [25]. Corneas were immersed in a closed vial of 100 ml of a commercial medium containing 2% fetal calf serum (FCS; CorneaMax, Eurobio, Les Ulis, France, http://www.eurobio.fr) at 31 C, renewed after 2 weeks.…”

Section: Cornea Characteristicsmentioning

confidence: 99%

Revisited Microanatomy of the Corneal Endothelial Periphery: New Evidence for Continuous Centripetal Migration of Endothelial Cells in Humans

et al. 2012

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The control of corneal transparency depends on the integrity of its endothelial monolayer, which is considered nonregenerative in adult humans. In pathological situations, endothelial cell (EC) loss, not offset by mitosis, can lead to irreversible corneal edema and blindness. However, the hypothesis of a slow, clinically insufficient regeneration starting from the corneal periphery remains debatable. The authors have re-evaluated the microanatomy of the endothelium in order to identify structures likely to support this homeostasis model. Whole endothelia of 88 human corneas (not stored, and stored in organ culture) with mean donor age of 80 6 12 years were analyzed using an original flatmounting technique. In 61% of corneas, cells located at the extreme periphery (last 200 lm of the endothelium) were organized in small clusters with two to three cell layers around Hassall-Henle bodies. In 68% of corneas, peripheral ECs formed centripetal rows 830 6 295 lm long, with Descemet membrane furrows visible by scanning electron microscopy. EC density was significantly higher in zones with cell rows. When immunostained, ECs in the extreme periphery exhibited lesser differentiation (ZO-1, Actin, Na/ K ATPase, CoxIV) than ECs in the center of the cornea but preferentially expressed stem cell markers (Nestin, Telomerase, and occasionally breast cancer resistance protein) and, in rare cases, the proliferation marker Ki67. Stored corneas had fewer cell clusters but more Ki67-positive ECs. We identified a novel anatomic organization in the periphery of the human corneal endothelium, suggesting a continuous slow centripetal migration, throughout life, of ECs from specific niches.

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“…intraocular and eyelid pressure and eye rubbing (29)(30)(31). Dextran is a complex polysaccharide routinely used at a concentration of 5% to act as a deswelling agent during organ culture to preserve human corneas (32)(33)(34)(35) although to date its use in controlling swelling during decellularization has not been investigated. Dextran increases colloidal osmotic pressure in cornea storage medium, limiting movement of water into the stroma by maintaining a high osmotic pressure (34).…”

Section: Introductionmentioning

confidence: 99%