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Karyotype changes in cultured human corneal endothelial cells

PURPOSE: To examine karyotype changes in cultured human corneal endothelial cells (HCECs). METHODS: HCECs with Descemet’s membrane were removed from 20 donors of various ages (range, 2–77 years; average, 43.7±26.4 years) and cultured on dishes coated with extracellular matrix produced by bovine corn...

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Autores principales: Miyai, Takashi, Maruyama, Yoko, Osakabe, Yasuhiro, Nejima, Ryohei, Miyata, Kazunori, Amano, Shiro
Formato: Texto
Lenguaje:English
Publicado: Molecular Vision 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2391083/
https://www.ncbi.nlm.nih.gov/pubmed/18509550
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author Miyai, Takashi
Maruyama, Yoko
Osakabe, Yasuhiro
Nejima, Ryohei
Miyata, Kazunori
Amano, Shiro
author_facet Miyai, Takashi
Maruyama, Yoko
Osakabe, Yasuhiro
Nejima, Ryohei
Miyata, Kazunori
Amano, Shiro
author_sort Miyai, Takashi
collection PubMed
description PURPOSE: To examine karyotype changes in cultured human corneal endothelial cells (HCECs). METHODS: HCECs with Descemet’s membrane were removed from 20 donors of various ages (range, 2–77 years; average, 43.7±26.4 years) and cultured on dishes coated with extracellular matrix produced by bovine corneal endothelial cells (BCECs). Karyotype changes were examined by G-band karyotyping of HCECs at the third passage from 12 donors and the fifth passage from 16 donors. The number of chromosomes was analyzed in 40–50 cells from the third and fifth passages of each HCEC preparation. A detailed karyotype analysis of 10–16 cells from the third and fifth passages of each HCEC preparation was performed. The frequency of aneuploid cells per case (the number of abnormal cells divided by the total number of cells examined at metaphase) was tested for correlation with age by Spearman’s correlation analysis. RESULTS: At the third passage, five cases (41.7%) showed an almost normal karyotype, and five cases (41.7%) showed sex chromosome loss. One case (8.3%) showed chromosome 21 trisomy. At the fifth passage, five cases (31.3%) showed an almost normal karyotype, and four cases (25%) showed sex chromosome loss. Three cases (18.8%) showed chromosome 8 trisomy, and one case (6.3%) showed chromosome 21 trisomy. Donor age and the frequency of aneuploidy had a statistically significant correlation at the fifth passage (R=0.653, p=0.042). CONCLUSIONS: Donor age and the frequency of aneuploidy have a positive correlation in cultured HCECs at the fifth passage. Therefore, HCECs for clinical therapies should be obtained from donors as young as possible. Karyotyping cultured HCECs is crucial before clinical application.
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spelling pubmed-23910832008-05-27 Karyotype changes in cultured human corneal endothelial cells Miyai, Takashi Maruyama, Yoko Osakabe, Yasuhiro Nejima, Ryohei Miyata, Kazunori Amano, Shiro Mol Vis Research Article PURPOSE: To examine karyotype changes in cultured human corneal endothelial cells (HCECs). METHODS: HCECs with Descemet’s membrane were removed from 20 donors of various ages (range, 2–77 years; average, 43.7±26.4 years) and cultured on dishes coated with extracellular matrix produced by bovine corneal endothelial cells (BCECs). Karyotype changes were examined by G-band karyotyping of HCECs at the third passage from 12 donors and the fifth passage from 16 donors. The number of chromosomes was analyzed in 40–50 cells from the third and fifth passages of each HCEC preparation. A detailed karyotype analysis of 10–16 cells from the third and fifth passages of each HCEC preparation was performed. The frequency of aneuploid cells per case (the number of abnormal cells divided by the total number of cells examined at metaphase) was tested for correlation with age by Spearman’s correlation analysis. RESULTS: At the third passage, five cases (41.7%) showed an almost normal karyotype, and five cases (41.7%) showed sex chromosome loss. One case (8.3%) showed chromosome 21 trisomy. At the fifth passage, five cases (31.3%) showed an almost normal karyotype, and four cases (25%) showed sex chromosome loss. Three cases (18.8%) showed chromosome 8 trisomy, and one case (6.3%) showed chromosome 21 trisomy. Donor age and the frequency of aneuploidy had a statistically significant correlation at the fifth passage (R=0.653, p=0.042). CONCLUSIONS: Donor age and the frequency of aneuploidy have a positive correlation in cultured HCECs at the fifth passage. Therefore, HCECs for clinical therapies should be obtained from donors as young as possible. Karyotyping cultured HCECs is crucial before clinical application. Molecular Vision 2008-05-19 /pmc/articles/PMC2391083/ /pubmed/18509550 Text en http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Miyai, Takashi
Maruyama, Yoko
Osakabe, Yasuhiro
Nejima, Ryohei
Miyata, Kazunori
Amano, Shiro
Karyotype changes in cultured human corneal endothelial cells
title Karyotype changes in cultured human corneal endothelial cells
title_full Karyotype changes in cultured human corneal endothelial cells
title_fullStr Karyotype changes in cultured human corneal endothelial cells
title_full_unstemmed Karyotype changes in cultured human corneal endothelial cells
title_short Karyotype changes in cultured human corneal endothelial cells
title_sort karyotype changes in cultured human corneal endothelial cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2391083/
https://www.ncbi.nlm.nih.gov/pubmed/18509550
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