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

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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.

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Abbreviations

HDFs:

Human dermal fibroblasts

IPSC:

Induced pluripotent stem cell

ECM:

Extracellular matrix

LMA:

Low melting point agarose

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Acknowledgments

This study was supported by Cellular and Molecular Research Center of Shahrekord University of Medical Sciences, Shahrekord, Iran (Grant No. 91-10-5). The authors would like to express their deepest gratitude to the staffs of Shahrekord Omid Nuclear Medicine Center for sincere cooperation.

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Authors and Affiliations

  1. Cellular and Molecular Research Center, Shahrekord University of Medical Sciences, Shahrekord, 8813833435, Iran

    Mehdi Allahbakhshian-Farsani, Narges Abdian, Payam Ghasemi-Dehkordi, Marzieh Sadeghiani, Morteza Hashemzadeh-Chaleshtori & Somayeh Khosravi-Farsani

  2. Clinical Biochemistry Research Center, Shahrekord University of Medical Sciences, Shahrekord, Iran

    Javad Saffari-Chaleshtori

  3. Department of Hematology and Blood Banking, Faculty of Allied Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Mehdi Allahbakhshian-Farsani

Authors
  1. Mehdi Allahbakhshian-Farsani
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  2. Narges Abdian
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  3. Payam Ghasemi-Dehkordi
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  4. Marzieh Sadeghiani
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  5. Javad Saffari-Chaleshtori
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  6. Morteza Hashemzadeh-Chaleshtori
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  7. Somayeh Khosravi-Farsani
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Correspondence to Morteza Hashemzadeh-Chaleshtori.

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Allahbakhshian-Farsani, M., Abdian, N., Ghasemi-Dehkordi, P. et al. Cytogentic analysis of human dermal fibroblasts (HDFs) in early and late passages using both karyotyping and comet assay techniques. Cytotechnology 66, 815–822 (2014). https://doi.org/10.1007/s10616-013-9630-y

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  • DOI: https://doi.org/10.1007/s10616-013-9630-y

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