title: [Characterization of epithelial primary culture from human conjunctiva].
abstract: OBJECTIVE
  To evaluate primary cultures from human conjunctiva supplemented with fetal bovine serum, autologous serum, and platelet-rich autologous serum, over human amniotic membrane and lens anterior capsules.
  
  
  METHODS
  One-hundred and forty-eight human conjunctiva explants were cultured in CnT50(®) supplemented with 1, 2.5, 5 and 10% fetal bovine serum, autologous serum and platelet-rich autologous serum. Conjunctival samples were incubated at 37°C, 5% CO2 and 95% HR, for 3 weeks.
  
  
  RESULTS
  The typical phenotype corresponding to conjunctival epithelial cells was present in all primary cultures. Conjunctival cultures had MUC5AC-positive secretory cells, K19-positive conjunctival cells, and MUC4-positive non-secretory conjunctival cells, but were not corneal phenotype (cytokeratin K3-negative) and fibroblasts (CD90-negative).
  
  
  CONCLUSIONS
  Conjunctiva epithelial progenitor cells were preserved in all cultures; thus, a cell culture in CnT50(®) supplemented with 1 to 5% autologous serum over human amniotic membrane can provide better information of epithelial cell differentiation for the conjunctival surface reconstruction.
authors.affiliation: Servicio de Oftalmología, Hospital Universitario Ramón y Cajal, Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, España. Electronic address: lrivas53@gmail.com.
authors.forename: L
authors.initials: L
authors.lastname: Rivas
authors.affiliation: Laboratorio de Ingeniería Celular, Hospital Universitario La Paz, Madrid, España.
authors.forename: A
authors.initials: A
authors.lastname: Blázquez
authors.affiliation: Servicio de Oftalmología, Hospital Universitario Ramón y Cajal, Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, España.
authors.forename: F J
authors.initials: FJ
authors.lastname: Muñoz-Negrete
authors.affiliation: Servicio de Oftalmología, Hospital Central de la Cruz Roja, Madrid, España.
authors.forename: S
authors.initials: S
authors.lastname: López
authors.affiliation: Servicio de Oftalmología, Hospital Universitario Ramón y Cajal, Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, España.
authors.forename: G
authors.initials: G
authors.lastname: Rebolleda
authors.affiliation: Laboratorio de Ingeniería Celular, Hospital Universitario La Paz, Madrid, España.
authors.forename: F
authors.initials: F
authors.lastname: Domínguez
authors.affiliation: Servicio de Oftalmología, Hospital Universitario La Paz, Madrid, España.
authors.forename: A
authors.initials: A
authors.lastname: Pérez-Esteban
mesh_terms: D000368:Aged; D000650:Amnion; D000818:Animals; D057866:Anterior Capsule of the Lens; D002417:Cattle; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D018920:Coculture Techniques; D003228:Conjunctiva / Q000166:cytology*; D003470:Culture Media; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology*; D005260:Female; D005312:Fetal Blood; D006801:Humans; D053539:Keratin-19 / Q000032:analysis; D008297:Male; D008875:Middle Aged; D055271:Mucin 5AC / Q000032:analysis; D055264:Mucin-4 / Q000032:analysis; D061251:Primary Cell Culture / Q000379:methods*; D044967:Serum; D013234:Stem Cells / Q000166:cytology
chemical_list: D003470:Culture Media; D053539:Keratin-19; C524658:MUC4 protein, human; C524678:MUC5AC protein, human; D055271:Mucin 5AC; D055264:Mucin-4
keywords: Autologous serum; Cell differentiation; Conjunctival epithelial cells; Cultivo primario; Células epiteliales conjuntivales; Diferenciación celular; Human amniotic membrane; Membrana amniótica humana; Primary culture; Suero autólogo