Comparison of the enzymatic and explant methods for the culture of keratinocytes isolated from human foreskin

  • Authors:
    • Mahmoud Orazizadeh
    • Mahmoud Hashemitabar
    • Somayeh Bahramzadeh
    • Freshteh Nejad Dehbashi
    • Sadegh Saremy
  • View Affiliations

  • Published online on: March 9, 2015     https://doi.org/10.3892/br.2015.442
  • Pages: 304-308
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Abstract

Currently, culture and growth keratinocytes are important stages in achieving a reliable and reproducible skin tissue. In the present study, two different methods, enzymatic and explant methods, for keratinocytes isolation from human foreskin were compared. Foreskins were cut into 2‑3 mm pieces and placed in trypsin at 4˚C overnight for separation of the epidermis from the dermis. Subsequently, these samples were divided into two groups: i) Keratinocytes separated from the epidermis by trypsin and ii) by the explant method. These keratinocytes were divided into two groups: i) With no feeder layer and ii) onto a type I collagen scaffold. The cells were evaluated using immunocytochemistry and 4',6‑diamidine‑2'‑phenylindole dihydrochloride (DAPI) staining. In the enzymatic treatment, after 7‑10 days no attached cells were found in the cell culture dishes. In the explant method, keratinocytes were separated after ~24 h, attached rapidly and formed big colonies into a collagen scaffold. In the absence of a feeder layer, small colonies were developed with rapid loss of proliferation within 2‑3 days. Keratinocytes showed positive immunoreactivity for the pan‑cytokeratin marker and keratinocytes' nuclei were clearly observed. This method could be applied and developed as a component of skin substitutes to treat burns and wounds and also in laboratory testing.
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May-2015
Volume 3 Issue 3

Print ISSN: 2049-9434
Online ISSN:2049-9442

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Spandidos Publications style
Orazizadeh M, Hashemitabar M, Bahramzadeh S, Dehbashi FN and Saremy S: Comparison of the enzymatic and explant methods for the culture of keratinocytes isolated from human foreskin. Biomed Rep 3: 304-308, 2015
APA
Orazizadeh, M., Hashemitabar, M., Bahramzadeh, S., Dehbashi, F.N., & Saremy, S. (2015). Comparison of the enzymatic and explant methods for the culture of keratinocytes isolated from human foreskin. Biomedical Reports, 3, 304-308. https://doi.org/10.3892/br.2015.442
MLA
Orazizadeh, M., Hashemitabar, M., Bahramzadeh, S., Dehbashi, F. N., Saremy, S."Comparison of the enzymatic and explant methods for the culture of keratinocytes isolated from human foreskin". Biomedical Reports 3.3 (2015): 304-308.
Chicago
Orazizadeh, M., Hashemitabar, M., Bahramzadeh, S., Dehbashi, F. N., Saremy, S."Comparison of the enzymatic and explant methods for the culture of keratinocytes isolated from human foreskin". Biomedical Reports 3, no. 3 (2015): 304-308. https://doi.org/10.3892/br.2015.442