title: Use of human serum for human corneal endothelial cell culture.
abstract: OBJECTIVE
  To study human corneal endothelial cells (HCECs) cultured in vitro with human serum (HS) supplemented media (HS-SM) compared with HCEC cultured in fetal bovine serum (FBS) supplemented media (FBS-SM).
  
  
  METHODS
  One cornea from a donor aged 21 years and a pair of corneas from a 16 year-old donor were obtained from the eye bank and used to create two different cell populations. At the first passage, the cell populations were equally divided and seeded in two different wells containing FBS-SM or HS-SM. In subsequent passages, HS-SM was compared with FBS-SM by morphology, growth curves, immunohistochemistry and real-time reverse-transcriptase PCR for endothelial cell markers.
  
  
  RESULTS
  No difference in morphology could be seen in P2, P5 or in any other passages for cells grown in the two media. By growth curves, cell counts were similar in FBS-SM and HS-SM from days 1 to 5, with a trend towards higher cell counts in HS-SM at day 7. Cells grown in FBS-SM and HS-SM media showed similar expression of endothelial cell markers when assessed by immunohistochemistry and real-time reverse-transcriptase PCR.
  
  
  CONCLUSIONS
  HS-SM was similar to FBS-SM for HCEC culture when assessed by cell morphology, proliferation and protein/gene expression.
authors.affiliation: Department of Ophthalmology, Cornea & Anterior Segment Service, Wilmer Eye Institute, Johns Hopkins School of Medicine, Baltimore, Maryland, USA Department of Ophthalmology, Federal University of São Paulo/Paulista School of Medicine, São Paulo, Brazil State University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Lucas Monferrari Monteiro
authors.initials: LM
authors.lastname: Vianna
authors.affiliation: Department of Ophthalmology, Cornea & Anterior Segment Service, Wilmer Eye Institute, Johns Hopkins School of Medicine, Baltimore, Maryland, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Kallay
authors.affiliation: Department of Ophthalmology, Cornea & Anterior Segment Service, Wilmer Eye Institute, Johns Hopkins School of Medicine, Baltimore, Maryland, USA.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Toyono
authors.affiliation: Department of Ophthalmology, Federal University of São Paulo/Paulista School of Medicine, São Paulo, Brazil.
authors.forename: Rubens
authors.initials: R
authors.lastname: Belfort
authors.affiliation: Lions VisionGift, Portland, Oregon, USA.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Holiman
authors.affiliation: Department of Ophthalmology, Cornea & Anterior Segment Service, Wilmer Eye Institute, Johns Hopkins School of Medicine, Baltimore, Maryland, USA.
authors.forename: Albert S
authors.initials: AS
authors.lastname: Jun
mesh_terms: D000293:Adolescent; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media; D004728:Endothelium, Corneal / Q000166:cytology* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction; D044967:Serum; D014019:Tissue Donors; D055815:Young Adult
chemical_list: D015415:Biomarkers; D003470:Culture Media
keywords: Cornea; Experimental &#8211 laboratory