Repository of Research and Investigative Information

Repository of Research and Investigative Information

Kurdistan University of Medical Sciences

Isolation, cultivation and transfection of human keratinocytes

(2014) Isolation, cultivation and transfection of human keratinocytes. Cell Biology International.

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Human keratinocytes could be used in the repair of damaged skin, in tissue engineering applications, gene therapy and recently, the generation of iPS cells. We isolated human keratinocytes from foreskin and subsequently cultured them on fibronectin, collagen type I, gelatin and laminin-coated dishes that contained three different types of serum-free medium (epilife, KSM or CnT). We developed improved conditions for efficient transfection of these human keratinocytes by testing three common transfection methods and a GFP plasmid vector. The isolated cells showed typical keratinocyte morphology and expressed the epithelial cell specific antigen, cytokeratin 14. Collagen type 1, epilife medium and lipofectamin 2000 gave the best results for isolation and transfection of human keratinocytes. Our protocol can be used as a reproducible, simple and efficient method for isolation, cultivation and genetic manipulation of human keratinocytes, which may be useful in cell and gene therapy applications.© 2013 International Federation for Cell Biology.

Item Type: Article
Keywords: collagen type 1; cytokeratin 14; fibronectin; gelatin; green fluorescent protein; laminin; lipofectamine; plasmid vector, antigen expression; article; cell adhesion; cell culture; cell growth; cell isolation; cell structure; controlled study; genetic transfection; human; human cell; immunofluorescence; keratinocyte; newborn, Ips, effectene; human keratinocyte; lipofectamine 2000; serum free medium; transfection, Antigens, Surface; Cell Proliferation; Cell Separation; Cells, Cultured; Collagen Type I; Culture Media; Fibronectins; Foreskin; Green Fluorescent Proteins; Humans; Keratin-14; Keratinocytes; Lipids; Male; Transfection
Page Range: pp. 444-451
Journal or Publication Title: Cell Biology International
Volume: 38
Number: 4
Publisher: Portland Press Ltd
Identification Number: 10.1002/cbin.10218
ISSN: 10656995
Depositing User: مهندس جمال محمودپور

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