Comparison of the effects between animal-derived trypsin and recombinant trypsin on human skin cells proliferation, gene and protein expression

Maarof Manira, Khairoji Khairul Anuar, Wan Tai Seet, Abd Wahab Ahmad Irfan, Min Hwei Ng, Chua Kien Hui, Mohd Heikal Mohd Yunus, Bin Saim Aminuddin, Ruszymah Idrus

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Animal-derivative free reagents are preferred in skin cell culture for clinical applications. The aim of this study was to compare the performance and effects between animal-derived trypsin and recombinant trypsin for skin cells culture and expansion. Full thickness human skin was digested in 0.6 % collagenase for 6 h to liberate the fibroblasts, followed by treatment with either animal-derived trypsin; Trypsin EDTA (TE) or recombinant trypsin; TrypLE Select (TS) to liberate the keratinocytes. Both keratinocytes and fibroblasts were then culture-expanded until passage 2. Trypsinization for both cell types during culture-expansion was performed using either TE or TS. Total cells yield was determined using a haemocytometer. Expression of collagen type I, collagen type III (Col-III), cytokeratin 10, and cytokeratin 14 genes were quantified via RT-PCR and further confirmed with immunocytochemical staining. The results of our study showed that the total cell yield for both keratinocytes and fibroblasts treated with TE or TS were comparable. RT-PCR showed that expression of skin-specific genes except Col-III was higher in the TS treated group compared to that in the TE group. Expression of proteins specific to the two cell types were confirmed by immunocytochemical staining in both TE and TS groups. In conclusion, the performance of the recombinant trypsin is comparable with the well-established animal-derived trypsin for human skin cell culture expansion in terms of cell yield and expression of specific cellular markers.

Original languageEnglish
Pages (from-to)41-49
Number of pages9
JournalCell and Tissue Banking
Volume15
Issue number1
DOIs
Publication statusPublished - 2014

Fingerprint

Ethylenediaminetetraacetic acid
Cell proliferation
Trypsin
Skin
Animals
Cell culture
Genes
Cell Proliferation
Proteins
Gene Expression
Fibroblasts
Collagen
Edetic Acid
Cell Culture Techniques
Keratinocytes
Collagen Type III
Derivatives
Keratin-14
Staining and Labeling
Polymerase Chain Reaction

Keywords

  • Animal-derived trypsin
  • Fibroblasts
  • Keratinocytes
  • Recombinant trypsin
  • Tissue engineering

ASJC Scopus subject areas

  • Transplantation
  • Biomaterials
  • Biomedical Engineering
  • Cell Biology
  • Medicine(all)

Cite this

Comparison of the effects between animal-derived trypsin and recombinant trypsin on human skin cells proliferation, gene and protein expression. / Manira, Maarof; Khairul Anuar, Khairoji; Seet, Wan Tai; Ahmad Irfan, Abd Wahab; Ng, Min Hwei; Kien Hui, Chua; Mohd Yunus, Mohd Heikal; Aminuddin, Bin Saim; Idrus, Ruszymah.

In: Cell and Tissue Banking, Vol. 15, No. 1, 2014, p. 41-49.

Research output: Contribution to journalArticle

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