title: A technique to harvest Descemet's membrane with viable endothelial cells for selective transplantation.
abstract: OBJECTIVE
  To describe a surgical technique using an artificial anterior chamber to facilitate harvest of Descemet's membrane (DM) and endothelium for corneal endothelial cell transplantation.
  
  
  METHODS
  Laboratory investigation.
  
  
  METHODS
  Corneoscleral buttons of seven human donor eyes were mounted endothelial side up on an artificial anterior chamber. Keeping the endothelial side with its usual concavity, a manual trephination was made on the posterior surface with a 9.0-mm trephine, inside the Schwalbe line and just past the DM in depth. The chamber was filled with air, causing the endothelial side of the donor cornea to assume a convex configuration. The DM along with its endothelium was separated from the posterior stroma using a blunt cyclodialysis spatula. Drops of trypan blue 0.3% and alizarin red S 0.2% (n = 6) were applied. The stained DMs were examined under a light microscope and photographed to calculate the percentage of endothelial cell damage. Histology was done on the unstained cornea.
  
  
  RESULTS
  The DM carrying endothelium was successfully removed from the posterior stroma in all seven eyes. Although the DM appears to be very friable, all samples were removed in toto without rupture. Vital staining showed a mean endothelial cell loss of 8.46% (standard deviation (SD) 6.9). Direct light microscopy demonstrated the preservation of endothelial cell morphology.
  
  
  CONCLUSIONS
  This technique appears to be a safe and straightforward method to harvest DM for endothelial cell transplantation. Further studies are underway to determine the optimal method of insertion of the obtained healthy DM with endothelial cells through small corneal incisions.
authors.affiliation: Department of Ophthalmology, University of California, Irvine, California, USA.
authors.forename: Teresa S
authors.initials: TS
authors.lastname: Ignacio
authors.forename: Thao T B
authors.initials: TT
authors.lastname: Nguyen
authors.forename: Melvin A
authors.initials: MA
authors.lastname: Sarayba
authors.forename: Paula M
authors.initials: PM
authors.lastname: Sweet
authors.forename: Omar
authors.initials: O
authors.lastname: Piovanetti
authors.forename: Roy S
authors.initials: RS
authors.lastname: Chuck
authors.forename: Ashley
authors.initials: A
authors.lastname: Behrens
mesh_terms: D000293:Adolescent; D000368:Aged; D000880:Anthraquinones; D018929:Cell Culture Techniques / Q000379:methods; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D017690:Cell Transplantation; D004396:Coloring Agents; D016039:Corneal Transplantation / Q000379:methods; D003886:Descemet Membrane / Q000601:surgery*; D004728:Endothelium, Corneal / Q000166:cytology* / Q000637:transplantation*; D006801:Humans; D008875:Middle Aged; D009885:Ophthalmology / Q000379:methods; D013194:Staining and Labeling / Q000379:methods; D014019:Tissue Donors; D014343:Trypan Blue
chemical_list: D000880:Anthraquinones; D004396:Coloring Agents; C004468:Alizarin Red S; D014343:Trypan Blue