title: Novel Identity and Functional Markers for Human Corneal Endothelial Cells.
abstract: Human corneal endothelial cell (HCEC) density decreases with age, surgical complications, or disease, leading to vision impairment. Such endothelial dysfunction is an indication for corneal transplantation, although there is a worldwide shortage of transplant-grade tissue. To overcome the current poor donor availability, here we isolate, expand, and characterize HCECs in vitro as a step toward cell therapy.
  
  
  
  Human corneal endothelial cells were isolated from cadaveric corneas and expanded in vitro. Cell identity was evaluated based on morphology and immunocytochemistry, and gene expression analysis and flow cytometry were used to identify novel HCEC-specific markers. The functional ability of HCEC to form barriers was assessed by transendothelial electrical resistance (TEER) assays.
  
  
  
  Cultured HCECs demonstrated canonical morphology for up to four passages and later underwent endothelial-to-mesenchymal transition (EnMT). Quality of donor tissue influenced cell measures in culture including proliferation rate. Cultured HCECs expressed identity markers, and microarray analysis revealed novel endothelial-specific markers that were validated by flow cytometry. Finally, canonical HCECs expressed higher levels of CD56, which correlated with higher TEER than fibroblastic HCECs.
  
  
  
  In vitro expansion of HCECs from cadaveric donor corneas yields functional cells identifiable by morphology and a panel of novel markers. Markers described correlated with function in culture, suggesting a basis for cell therapy for corneal endothelial dysfunction.
authors.affiliation: Shiley Eye Institute, University of California San Diego, La Jolla, California, United States.
authors.forename: Alena
authors.initials: A
authors.lastname: Bartakova
authors.affiliation: Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, Florida, United States.
authors.forename: Karen
authors.initials: K
authors.lastname: Alvarez-Delfin
authors.affiliation: Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, Florida, United States.
authors.forename: Alejandra D
authors.initials: AD
authors.lastname: Weisman
authors.affiliation: Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, Florida, United States.
authors.forename: Enrique
authors.initials: E
authors.lastname: Salero
authors.affiliation: Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, Florida, United States.
authors.forename: Gabriella A
authors.initials: GA
authors.lastname: Raffa
authors.affiliation: Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, Florida, United States.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Merkhofer
authors.affiliation: Shiley Eye Institute, University of California San Diego, La Jolla, California, United States 2Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, Florida, United States 3Emmecell, K.
authors.forename: Noelia J
authors.initials: NJ
authors.lastname: Kunzevitzky
authors.affiliation: Shiley Eye Institute, University of California San Diego, La Jolla, California, United States 2Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, Florida, United States 4Byers Eye I.
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Goldberg
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000378:metabolism*; D002102:Cadaver; D002452:Cell Count; D049109:Cell Proliferation; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003316:Corneal Diseases / Q000378:metabolism* / Q000473:pathology / Q000601:surgery; D016039:Corneal Transplantation; D017097:Electric Impedance; D004728:Endothelium, Corneal / Q000378:metabolism* / Q000473:pathology; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D008875:Middle Aged; D014019:Tissue Donors; D055815:Young Adult
chemical_list: D015415:Biomarkers