Lucas Monferrari Monteiro Vianna1, Laura Kallay2, Tetsuya Toyono2, Rubens Belfort3, Jeffrey D Holiman4, Albert S Jun2. 1. 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. 2. Department of Ophthalmology, Cornea & Anterior Segment Service, Wilmer Eye Institute, Johns Hopkins School of Medicine, Baltimore, Maryland, USA. 3. Department of Ophthalmology, Federal University of São Paulo/Paulista School of Medicine, São Paulo, Brazil. 4. Lions VisionGift, Portland, Oregon, USA.
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. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.
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. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.
Authors: Bart Kramer; Claudio Corallo; Angelique van den Heuvel; Justin Crawford; Thomas Olivier; Edo Elstak; Nicola Giordano; Paul Vulto; Henriette L Lanz; Richard A J Janssen; Michela A Tessari Journal: Sci Rep Date: 2022-10-08 Impact factor: 4.996
Authors: Gary S L Peh; Heng-Pei Ang; Chan N Lwin; Khadijah Adnan; Benjamin L George; Xin-Yi Seah; Shu-Jun Lin; Maninder Bhogal; Yu-Chi Liu; Donald T Tan; Jodhbir S Mehta Journal: Sci Rep Date: 2017-10-26 Impact factor: 4.379