Cytogentic analysis of human dermal fibroblasts (HDFs) in early and late passages using both karyotyping and comet assay techniques

Allahbakhshian-Farsani, M. and Abdian, N. and Ghasemi-Dehkordi, P. and Sadeghiani, M. and Saffari-Chaleshtori, J. and Hashemzadeh-Chaleshtori, M. and Khosravi-Farsani, S. (2014) Cytogentic analysis of human dermal fibroblasts (HDFs) in early and late passages using both karyotyping and comet assay techniques. CYTOTECHNOLOGY, 66 (5). pp. 815-822.

[img]
Preview
Text
31.pdf

Download (570kB) | Preview

Abstract

Human dermal fibroblasts (HDFs) are a potential source of somatic cells for genetic manipulation and tissue engineering. Confirmation of cytogenetic stability of these cells is an essential step for cell nuclear transfer and generation of a suitable and functional induced pluripotent stem cells line. HDF cells were isolated and cultured from human foreskin samples. Cytogenetic stability of these cells was evaluated in early (3-4) and late (10-15) passages using karyotype test and alkaline comet assay techniques. HDF cells in early and late passages showed normal karyotype but by comet assay abnormality and DNA damages in late passages of HDFs were observed. Also, the parameters of alkaline comet assay in early passages of HDFs compared with late passages and positive control groups more significantly were different (p < 0.05). These findings indicate that single-strand breaks or DNA damage after many passages may have occurred in HDF cells. Our results demonstrate that only early passages of HDF cells maintain cytogenetic stability and are good candidates for gene reprogramming. In conclusion, karyotype testing alone can not be used for detection of all signs of cytogenetic abnormality and DNA damages of cells. So, for precise evaluation of DNA damage and cytogenetic instability of fibroblast cells comet assay and karyotype techniques could complement each other.

Item Type: Article
Uncontrolled Keywords: HDF; Cytogenetic analysis; Karyotype test; Comet assay; IPSC
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: 30 Jul 2017 08:28
Last Modified: 30 Jul 2017 08:28
URI: http://eprints.skums.ac.ir/id/eprint/2062

Actions (login required)

View Item View Item