Purchase this article with an account.
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.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
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.
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.
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.
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.
This PDF is available to Subscribers Only