September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Anterior lens capsule for endothelial grafts production.
Author Affiliations & Notes
  • Carlos-Alberto Rodriguez-Barrientos
    Ophthalmology, Tecnologico de Monterrey, Monterrey, Nuevo Leon, Mexico
    Instituto Universitario Fernández-Vega, Oviedo, Spain
  • Alvaro Meana
    Instituto Universitario Fernández-Vega, Oviedo, Spain
  • Manuel Chacon
    Instituto Universitario Fernández-Vega, Oviedo, Spain
  • Natalia Vazquez
    Instituto Universitario Fernández-Vega, Oviedo, Spain
  • Judith Zavala
    Ophthalmology, Tecnologico de Monterrey, Monterrey, Nuevo Leon, Mexico
  • Jorge E Valdez
    Ophthalmology, Tecnologico de Monterrey, Monterrey, Nuevo Leon, Mexico
  • Jesus Merayo-Lloves
    Instituto Universitario Fernández-Vega, Oviedo, Spain
  • Footnotes
    Commercial Relationships   Carlos-Alberto Rodriguez-Barrientos, None; Alvaro Meana, None; Manuel Chacon, None; Natalia Vazquez, None; Judith Zavala, None; Jorge Valdez, None; Jesus Merayo-Lloves, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 5294. doi:
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      Carlos-Alberto Rodriguez-Barrientos, Alvaro Meana, Manuel Chacon, Natalia Vazquez, Judith Zavala, Jorge E Valdez, Jesus Merayo-Lloves; Anterior lens capsule for endothelial grafts production.. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5294.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : The objective of the present study is to investigate the use of anterior lens capsule (ALC) as scaffold for culture of human corneal endothelial cells (hCEC) for endothelial graft production.

Methods : 1-Donors: Fresh human lens and corneoescleral rims were obtained from our local tissue bank after informed consent.
2-Scaffold development: ALC was dissected under stereoscopic dissecting microscope and trephined using a 8mm diameter corneal punch. ALC were decellularized using trypsin/EDTA for 10 minutes and finally were crosslinked using 254nm UV irradiation lamp to improve their resistance.
3-Cellular culture: hCEC were obtained from normal human corneoscleral rims obtained during surgery after 8mm trephination of the graft. Peripheral Descemet membrane, along with endothelial cells, was dissected and seeded on a culture dish as an explant. Once confluent, endothelial cells were subcultured on an ALC decellularized scaffold.
4-Cellular analysis: Cell morphology analysis was studied by phase contrast microscopy and immunochemical analyses were performed against Na+K+/ATPase and ZO-1 markers.
5-Ex vivo experimental transplant: Cultured ALC were transplanted onto an anterior lamellar segment using an ex vivo bioreactor and cultured for 7 days. Following this, lamellar segments with ALC were fixed on ice-cold methanol, sectioned and stained using hematoxilin-eosin.

Results : ALC was effectively cultured with hCEC observing, under phase contrast microscopy, hCEC as a compact monolayer with regular polygonal morphology. Immunostaining analyses show the expression of ZO-1 and Na+K+/ATPase as characteristic markers for hCEC. Moreover, ALC with hCEC was completely integrated as a corneal endothelium on an ex vivo experimental transplant.

Conclusions : Using the remnant ocular tissues such as human lens and peripheral endothelial rims, a tissue engineered corneal endothelium could be achieved using ALC and hCEC.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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