A study of cytogenetic stability of induced pluripotent stem cells using karyotyping and comet assay techniques

Ghasemi-Dehkordi, Payam. and Allahbakhshian-Farsani, Mehdi. and Abdian, Narges. and Jafari-Ghahfarokhi, Hamideh. and Saffari-Chaleshtori, Javad. and Sadeghiani, Marzieh. and Mirzaeian, Amin. and Hashemzadeh-Chaleshtori, Morteza. (2015) A study of cytogenetic stability of induced pluripotent stem cells using karyotyping and comet assay techniques. Journal of Kerman University of Medical Sciences, 22 (4). pp. 370-384.

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Abstract

Background & Aims: Induced pluripotent stem cells (iPSCs) have the capability to undergo unlimited selfrenewal and differentiation into all cell types in the body. These cells are artificially derived from a nonpluripotent cell, typically human dermal fibroblasts (HDFs). The study of cytogenetic stability of these cells, in order to use iPS cells and apply studies in therapeutic applications, is essential. Methods: In the present experimental study, HDFs were isolated and cultured from human foreskin samples. The cytogenetic stability of these cells was evaluated in early passages (1-3) of HDFs using karyotype test and alkaline comet assay technique. The HDF cells treatment with hydrogen peroxide (H2O2) was used as a positive control for alkaline comet assay. The iPS cells with low passage (4-7) derived from reprogrammed HDFs were cultured on mouse embryonic fibroblast (MEF) feeder layer and cytogenetic stability of these cells were evaluated in early passages using karyotype test and alkaline comet assay technique. Results: The iPS cells in early passages (4-7) had normal karyotype (46, XY) and DNA damage and comet were not observed in these cells. In addition, HDF cells showed normal karyotype in early passages (1-3), but using comet assay, abnormality and DNA damages were observed in positive control (HDFs treated with H2O2). The comparison of alkaline comet assay parameters of iPS and HDF cells with positive control group showed statistically significant differences (P < 0.05). Conclusion: Since the comet assay is a sensitive technique for finding DNA damage, it is best if cytogenetic stability of these cells were evaluated before performing functional experiments on iPS cells. Therefore, for the precise evaluation of DNA damage and cytogenetic stability of iPS cells, the two techniques could complement each other. © 2015, Kerman University of Medical Sciences. All rights reserved.

Item Type: Article
Additional Information: cited By 0
Uncontrolled Keywords: hydrogen peroxide, Article; chromosome analysis; comet assay; controlled study; cytogenetics; DNA damage; fibroblast; human; karyotyping; mouse; nonhuman; pluripotent stem cell
Subjects: WR Dermatology
QU Biochemistry > Cell biology and genetics
Divisions: Reserach Vice-Chancellar Department > Cellular and Molecular Research Center
Reserach Vice-Chancellar Department > Clinical Biochemistry Research Center
Depositing User: zahra bagheri .
Date Deposited: 29 Jul 2017 05:37
Last Modified: 03 Mar 2018 08:05
URI: http://eprints.skums.ac.ir/id/eprint/1920

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