Storage of human fetal retina in optisol prior to subretinal transplantation

David A. Diloreto, Coca Del Cerro, Eliot S. Lazar, Christopher Cox, Manuel Del Cerro

Research output: Contribution to journalArticle

Abstract

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 of 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°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). Non-transplanted 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.

Original languageEnglish (US)
Pages (from-to)553-561
Number of pages9
JournalCell Transplantation
Volume5
Issue number5
DOIs
StatePublished - Sep 1996
Externally publishedYes

Fingerprint

Retina
Animals
Transplantation
Cells
Tissue
Sodium bicarbonate
Transplantation (surgical)
Dextran
Antioxidants
Grafts
Optical microscopy
Amino acids
Rats
Cell Survival
Sodium
HEPES
Light
Sodium Bicarbonate
Fluids
Chondroitin Sulfates

Keywords

  • Cell viability
  • Immunosuppression
  • Light damage
  • Retina transplantation
  • Storage medium

ASJC Scopus subject areas

  • Cell Biology
  • Transplantation

Cite this

Storage of human fetal retina in optisol prior to subretinal transplantation. / Diloreto, David A.; Del Cerro, Coca; Lazar, Eliot S.; Cox, Christopher; Del Cerro, Manuel.

In: Cell Transplantation, Vol. 5, No. 5, 09.1996, p. 553-561.

Research output: Contribution to journalArticle

Diloreto, David A. ; Del Cerro, Coca ; Lazar, Eliot S. ; Cox, Christopher ; Del Cerro, Manuel. / Storage of human fetal retina in optisol prior to subretinal transplantation. In: Cell Transplantation. 1996 ; Vol. 5, No. 5. pp. 553-561.
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abstract = "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 of 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°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). Non-transplanted 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.",
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