Serum-free primary human fibroblast and keratinocyte coculture

Tissue Engineering. Part a
Sally MujajSean Richards

Abstract

Research has shown that the inclusion of a fibroblast cell support layer is required for the isolation and expansion of primary keratinocytes. Recent advances have provided keratinocyte culture with fibroblast-free alternatives. However, these technologies are often undefined and rely on the incorporation of purified proteins/components. To address this problem we developed a medium that used recombinant proteins to support the serum-free isolation and expansion of human dermal fibroblasts and keratinocytes. The human dermal fibroblasts were able to be isolated serum free by adding recombinant human albumin to a collagenase solution. These fibroblasts were then expanded using a serum-free medium containing recombinant proteins: epidermal growth factor, basic fibroblast growth factor, chimeric vitronectin:insulin-like growth factor-I protein, and recombinant human albumin. These fibroblasts maintained a typical morphology and expressed fibroblast markers during their serum-free isolation, expansion, and freezing. Moreover, these fibroblasts were able to support the serum-free isolation and expansion of primary keratinocytes using these recombinant proteins. Real-time polymerase chain reaction and immunofluorescence analysis conf...Continue Reading

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Citations

Aug 2, 2012·Cell and Tissue Banking·J DragúňováJ Koller
Oct 20, 2010·Tissue Engineering. Part B, Reviews·Benoit HendrickxAernout Luttun
Dec 5, 2015·International Wound Journal·Meirong LiWeidong Han
Apr 10, 2015·Journal of Biomedical Materials Research. Part a·Omolbanin MirdailamiFatemeh Atyabi
Mar 16, 2013·Experimental Dermatology·Rebecca P HillColin A Jahoda
Feb 7, 2015·Tissue Engineering. Part B, Reviews·Sara LlamesMarcela del Río
Oct 16, 2015·Journal of Dental Research·G YuH Egusa
Jul 7, 2017·Cell Proliferation·Nazmul HaqueMohammad Tariqur Rahman

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Methods Mentioned

BETA
FCS
scraping
PCR

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