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Use of human serum for human corneal endothelial cell culture
  1. Lucas Monferrari Monteiro Vianna1,2,3,
  2. Laura Kallay1,
  3. Tetsuya Toyono1,
  4. Rubens Belfort Jr2,
  5. Jeffrey D Holiman4,
  6. Albert S Jun1
  1. 1Department of Ophthalmology, Cornea & Anterior Segment Service, Wilmer Eye Institute, Johns Hopkins School of Medicine, Baltimore, Maryland, USA
  2. 2Department of Ophthalmology, Federal University of São Paulo/Paulista School of Medicine, São Paulo, Brazil
  3. 3State University of Rio de Janeiro, Rio de Janeiro, Brazil
  4. 4Lions VisionGift, Portland, Oregon, USA
  1. Correspondence to Dr Albert S Jun, Department of Ophthalmology, Cornea & Anterior Segment Service, Wilmer Eye Institute, Johns Hopkins School of Medicine, 400 N. Broadway, Smith Building 5011, Baltimore, MD 21231, USA; aljun{at}jhmi.edu

Abstract

Background/aims 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.

  • Cornea
  • Experimental &#8211 laboratory

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