title: An Innovative Surgical Technique for Subretinal Transplantation of Human Embryonic Stem Cell-Derived Retinal Pigmented Epithelium in Yucatan Mini Pigs: Preliminary Results.
abstract: OBJECTIVE
  To develop a safe and efficient surgical procedure for subretinal implantation into porcine eyes of a human embryonic stem cell-derived retinal pigmented epithelium (hESC-RPE) monolayer seeded onto a Parylene-C scaffold. This implant is referred to as CPCB-RPE1.
  
  
  METHODS
  Ultrathin Parylene-C scaffolds were seeded with hESC-RPE and surgically implanted into the subretinal space of Yucatan mini pigs (n = 8). The surgery consisted of pars plana vitrectomy, induction of a limited retinal detachment, and peripheral retinotomy for insertion of the monolayer using a novel tissue injector, followed by silicone oil tamponade injection, laser photocoagulation around the retinotomy site, and inferior iridectomy. Oral cyclosporine was administered from day 1 and during the entire follow-up period. Three months later, the animals were euthanized and the eyes and major organs were submitted for histological analysis. Adjacent sections underwent immunohistochemical analysis to detect human cells using anti-TRA-1-85 (human blood group antigen) antibody and DAPI antibodies.
  
  
  RESULTS
  The cell monolayer was immunopositive for TRA-1-85 3 months after implantation and migration from the Parylene-C scaffold was not detected. One eye had a mild inflammatory reaction around the implant that was negative for human biomarkers. No intraocular or systemic tumors were detected.
  
  
  CONCLUSIONS
  The hESC-RPE cells survived for 3 months in this animal model. The surgical procedure for subretinal implantation of CPCB-RPE1 is feasible and safe, without cell migration off the scaffold or development of ocular or systemic tumors.
authors.forename: Rodrigo A
authors.initials: RA
authors.lastname: Brant Fernandes
authors.forename: Michael Janusz
authors.initials: MJ
authors.lastname: Koss
authors.forename: Paulo
authors.initials: P
authors.lastname: Falabella
authors.forename: Francisco Rosa
authors.initials: FR
authors.lastname: Stefanini
authors.forename: Mauricio
authors.initials: M
authors.lastname: Maia
authors.forename: Bruno
authors.initials: B
authors.lastname: Diniz
authors.forename: Ramiro
authors.initials: R
authors.lastname: Ribeiro
authors.forename: Yuntao
authors.initials: Y
authors.lastname: Hu
authors.forename: David
authors.initials: D
authors.lastname: Hinton
authors.forename: Dennis O
authors.initials: DO
authors.lastname: Clegg
authors.forename: Gerald
authors.initials: G
authors.lastname: Chader
authors.forename: Mark S
authors.initials: MS
authors.lastname: Humayun
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005451:Fluorescein Angiography; D000066449:Human Embryonic Stem Cells / Q000637:transplantation*; D006801:Humans; D013508:Ophthalmologic Surgical Procedures; D011108:Polymers; D012160:Retina / Q000000981:diagnostic imaging / Q000601:surgery*; D055213:Retinal Pigment Epithelium / Q000166:cytology*; D033581:Stem Cell Transplantation; D013552:Swine; D013556:Swine, Miniature; D054457:Tissue Scaffolds; D041623:Tomography, Optical Coherence; D014183:Transplantation, Heterologous; D014992:Xylenes
chemical_list: D011108:Polymers; D014992:Xylenes; C011055:parylene