Storage of human fetal retina in optisol prior to subretinal transplantation.
We have previously reported that Optisol, a commercially available medium (Chiron Vision, Irvine, CA), is a favorable fluid for the storage of neural retina. We now characterize the cell survival and histologic integrity of the tissue following storage and then transplantation. Neural retinas from human fetal eyes obtained within 1 h or pregnancy termination were stored in Optisol-GS medium (containing TC-199 and MEM medium, 2.5% chondroitin sulfate, 1% dextran, HEPES buffer, gentamicin sulfate, 0.1 mmol/L nonessential amino acids, sodium bicarbonate, 1 mmol/L sodium pyruvate, and additional antioxidants), at 4 degrees C for the following time periods: 0, 1, 2, and 7 days, and then tested for viability. Retinas stored for 2 and 7 days were mechanically dissociated and grafted into the eyes of light-damaged Fischer 344 rats (17 animals, 28 eyes). Nontransplanted eyes received injections of vehicle only (5 animals, 6 eyes). All animals were immunosuppressed daily with cyclosporine (10 mg/kg). All experiments were conducted in strict accordance with institutional, federal, and ARVO guidelines. Cell viability averaged 94.8%, 90.2%, 83.2%, and 76.8% at 0, 1, 2, and 7 days storage, respectively. Light microscopy demonstrated that the fetal retina was preserved without evident changes for up to 48 h. Up to 7 days there was a good to very good preservation of the cells in the outer neuroblastic layer. One month after transplantation subretinal grafts of retinal tissue stored for 2 and 7 days in Optisol showed good integration with host retina and initial photoreceptor differentiation. Optisol is a widely used medium for the preservation of human corneas prior to transplantation. Our results show that this medium is also highly suitable for preserving human fetal retinas prior to transplantation.
Department of Opthalmology, University of Rochester School of Medicine, NY 14642, USA.
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DiLoreto
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del Cerro
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Lazar
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Cox
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del Cerro
D000818:Animals; D002470:Cell Survival; D002809:Chondroitin Sulfates; D045424:Complex Mixtures; D016895:Culture Media, Serum-Free; D003911:Dextrans; D005260:Female; D005839:Gentamicins; D006801:Humans; D011247:Pregnancy; D051381:Rats; D011916:Rats, Inbred F344; D012160:Retina / Q000196:embryology / Q000637:transplantation*; D014021:Tissue Preservation / Q000379:methods*
D045424:Complex Mixtures; D016895:Culture Media, Serum-Free; D003911:Dextrans; D005839:Gentamicins; D002809:Chondroitin Sulfates; C075736:Optisol