title: Hyaluronan production and hyaluronan synthase expression in three human conjunctiva-derived cell strains.
abstract: OBJECTIVE
  The conjunctiva maintains the health of the ocular surface by protecting the eye from pathogen invasion, injury, and dryness. In this study, we investigated the regulation of hyaluronan (HA) synthesis by cytokines in conjunctiva-derived cells.
  
  
  METHODS
  Cultured primary cells derived from human conjunctivas that had been removed as surgical specimens were transfected with an immortalizing gene (human papilloma virus 16 E6/E7). To compare the biological features of the primary and immortalized cells, we assessed their morphological features and gene expression patterns. We also examined the effects of inflammatory cytokines on hyaluronan synthase (HAS) expression and HA production.
  
  
  RESULTS
  Three conjunctiva-derived cell strains were established and could be passaged up to 15 times. All strains expressed β2MG and KRT13 transcripts, highly expressed in conjunctival epithelial cells. HA production and expression of the three HAS isoforms were detected in the cell strains; however, cytokine treatment had no significant effect on HA production or HAS isoform expression.
  
  
  CONCLUSIONS
  We succeeded in deriving three human cell strains from conjunctival tissue. In the conjunctiva-derived cell strains, HA production and HAS mRNA expression were stable and were not changed by either TGF-β or PDGF-BB.
authors.affiliation: Department of Ophthalmology and Visual Sciences, Faculty of Medicine, Yamagata University, 2-2-2 Iida-nishi, Yamagata, Yamagata 990-9585, Japan. kasiwagi@med.id.yamagata-u.ac.jp
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Kashiwagi
authors.forename: Koichi
authors.initials: K
authors.lastname: Nishitsuka
authors.forename: Teiko
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Takamura
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Yamashita
mesh_terms: D000293:Adolescent; D000368:Aged; D000077214:Becaplermin; D002460:Cell Line; D002470:Cell Survival; D003228:Conjunctiva / Q000187:drug effects / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D014453:Glucuronosyltransferase / Q000235:genetics*; D006801:Humans; D000076002:Hyaluronan Synthases; D006820:Hyaluronic Acid / Q000096:biosynthesis*; D008297:Male; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
chemical_list: D020574:Proto-Oncogene Proteins c-sis; D012333:RNA, Messenger; D053773:Transforming Growth Factor beta1; D000077214:Becaplermin; D006820:Hyaluronic Acid; D014453:Glucuronosyltransferase; D000076002:Hyaluronan Synthases