title: Establishment of a human corneal epithelial cell line lacking the functional TACSTD2 gene as an in vitro model for gelatinous drop-like dystrophy.
abstract: OBJECTIVE
  Gelatinous drop-like corneal dystrophy (GDLD) is characterized by subepithelial amyloid deposition that engenders severe vision loss. The exact mechanism of this disease has yet to be elucidated. No fundamental treatment exists. This study was conducted to establish an immortalized corneal epithelial cell line to be used as a GDLD disease model.
  
  
  METHODS
  A corneal tissue specimen was obtained from a GDLD patient during surgery. Corneal epithelial cells were enzymatically separated from the cornea and were dissociated further into single cells. The epithelial cells were immortalized by the lentiviral transduction of the simian virus 40 (SV40) large T antigen and human telomerase reverse transcriptase (hTERT) genes. For the immortalized cells, proliferative kinetics, gene expressions, and functional analyses were performed.
  
  
  RESULTS
  The immortalized corneal epithelial cells continued to proliferate despite cumulative population doubling that exceeded 100. The cells showed almost no sign of senescence and displayed strong colony-forming activity. The cells exhibited a low epithelial barrier function as well as decreased expression of tight-junction-related proteins claudin 1 and 7. Using the immortalized corneal epithelial cells derived from a GDLD patient, we tested the possibility of gene therapy.
  
  
  CONCLUSIONS
  We established an immortalized corneal epithelial cell line from a GDLD patient. The immortalized cells exhibited cellular phenotypes similar to those of in vivo GDLD. The immortalized cells are thought to be useful for the development of new therapies for treating GDLD corneas and for elucidation of the pathophysiology of GDLD.
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Kitazawa
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kawasaki
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Shinomiya
authors.forename: Keita
authors.initials: K
authors.lastname: Aoi
authors.forename: Akira
authors.initials: A
authors.lastname: Matsuda
authors.forename: Toshinari
authors.initials: T
authors.lastname: Funaki
authors.forename: Kenta
authors.initials: K
authors.lastname: Yamasaki
authors.forename: Mina
authors.initials: M
authors.lastname: Nakatsukasa
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Ebihara
authors.forename: Akira
authors.initials: A
authors.lastname: Murakami
authors.forename: Junji
authors.initials: J
authors.lastname: Hamuro
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kinoshita
mesh_terms: D028226:Amyloidosis, Familial / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D003317:Corneal Dystrophies, Hereditary / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D019573:Epithelium, Corneal / Q000378:metabolism / Q000473:pathology*; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D008875:Middle Aged; D012313:RNA / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499380:TACSTD2 protein, human; D012313:RNA
keywords: corneal dystrophy; gelatinous drop-like; immortalized cells