Characterization of cryopreserved primary human corneal endothelial cells cultured in human serum-supplemented media

Characterization of cryopreserved primary human corneal endothelial cells cultured in human serum-supplemented media

Title: Characterization of cryopreserved primary human corneal endothelial cells cultured in human serum-supplemented media;
Caracterização de células humanas primárias criopreservados córnea endoteliais cultivadas em meio suplementado com soro humano
Author Vianna, Lucas Monferrari Monteiro [UNIFESP Google Scholar
Li, Hao-Dong Google Scholar
Holiman, Jeffrey D. Google Scholar
Stoeger, Christopher Google Scholar
Belfort, Rubens Junior Autor UNIFESP Google Scholar
Jun, Albert S. Google Scholar
Abstract Purpose: To compare cryopreserved human corneal endothelial cells (HCECs) grown in human serum-supplemented media (HS-SM) with cryopreserved HCECs grown in fetal bovine serum-supplemented media (FBS-SM). Methods: Three pairs of human corneas from donors aged 8, 28, and 31 years were obtained from the eye bank. From each pair, one cornea was used to start a HCEC culture using HS-SM

the other cornea was grown in FBS-SM. On reaching confluence, the six cell populations were frozen using 10% dimethyl sulfoxide-containing medium. Thawed cells grown in HS-SM were compared with those grown in FBS-SM with respect to morphology, growth curves, immunohistochemistry, real time-reverse transcriptase polymerase chain reaction (RT-PCR) for endothelial cell markers, and detachment time. Results: No difference in morphology was observed for cells grown in the two media before or after cryopreservation. By growth curves, cell counts after thawing were similar in both media, with a slight trend toward higher cell counts in FBS-SM. Cells grown in both the media demonstrated a similar expression of endothelial cell markers when assessed by immunohistochemistry, although HCEC marker gene expression was higher in cells grown in HS-SM than in those grown in FBS-SM as assessed by RT-PCR. With FBS-SM, there was a tendency of longer detachment time and lower cell passages. Conclusions: HS-SM was similar to FBS-SM for cryopreservation of cultured HCECs as assessed by analysis of cell morphology, proliferation, and protein expression, although marker gene expression was higher in cells grown in HS-SM than in those grown in FBS-SM. Detachment time was longer with FBS-SM and in lower passages.
Keywords Cryopreservation
Cornea
Endothelium
Corneal
Cell Culture Techniques
Culture MediaProliferative Capacity
Transplantation
Language English
Sponsor Brazilian government CAPES Foundation
State University of Rio de Janeiro
Date 2016
Published in Arquivos Brasileiros De Oftalmologia. Sao paulo, v. 79, n. 1, p. 37-41, 2016.
ISSN 0004-2749 (Sherpa/Romeo, impact factor)
Publisher Consel brasil oftalmologia
Extent 37-41
Origin http://dx.doi.org/10.5935 / 0004-2.749,20160011
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000369656700011
URI http://repositorio.unifesp.br/handle/11600/49610

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