Mesenchymal stem cell-conditioned medium accelerates skin wound healing: an in vitro study of fibroblast and keratinocyte scratch assays

Merlin Walter, Karina T. Wright, H.R. Fuller, Sheila MacNeil, W. Eustace B. Johnson

Research output: Contribution to journalArticle

Abstract

We have used in vitro scratch assays to examine the relative contribution of dermal fibroblasts and keratinocytes in the wound repair process and to test the influence of mesenchymal stem cell (MSC) secreted factors on both skin cell types. Scratch assays were established using single cell and co-cultures of L929 fibroblasts and HaCaT keratinocytes, with wound closure monitored via time-lapse microscopy. Both in serum supplemented and serum free conditions, wound closure was faster in L929 fibroblast than HaCaT keratinocyte scratch assays, and in co-culture the L929 fibroblasts lead the way in closing the scratches. MSC-CM generated under serum free conditions significantly enhanced the wound closure rate of both skin cell types separately and in co-culture, whereas conditioned medium from L929 or HaCaT cultures had no significant effect. This enhancement of wound closure in the presence of MSC-CM was due to accelerated cell migration rather than increased cell proliferation. A number of wound healing mediators were identified in MSC-CM, including TGF-beta1, the chemokines IL-6, IL-8, MCP-1 and RANTES, and collagen type I, fibronectin, SPARC and IGFBP-7. This study suggests that the trophic activity of MSC may play a role in skin wound closure by affecting both dermal fibroblast and keratinocyte migration, along with a contribution to the formation of extracellular matrix.
Original languageEnglish
Pages (from-to)1271-1281
Number of pages10
JournalExperimental Cell Research
Volume316
Issue number7
Early online date3 Mar 2010
DOIs
Publication statusPublished - 15 Apr 2010

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Conditioned Culture Medium
Mesenchymal Stromal Cells
Keratinocytes
Wound Healing
Fibroblasts
Skin
Wounds and Injuries
Coculture Techniques
Serum
Transforming Growth Factor beta1
Chemokine CCL5
Stem Cell Factor
Collagen Type I
Interleukin-8
Fibronectins
Chemokines
Cell Movement
Extracellular Matrix
In Vitro Techniques
Microscopy

Keywords

  • mesenchymal stem cells
  • wound healing
  • dermal fibroblast
  • keratinocyte
  • cell migration
  • secretome

Cite this

Walter, Merlin ; Wright, Karina T. ; Fuller, H.R. ; MacNeil, Sheila ; Johnson, W. Eustace B. / Mesenchymal stem cell-conditioned medium accelerates skin wound healing : an in vitro study of fibroblast and keratinocyte scratch assays. In: Experimental Cell Research. 2010 ; Vol. 316, No. 7. pp. 1271-1281.
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Mesenchymal stem cell-conditioned medium accelerates skin wound healing : an in vitro study of fibroblast and keratinocyte scratch assays. / Walter, Merlin; Wright, Karina T.; Fuller, H.R.; MacNeil, Sheila; Johnson, W. Eustace B.

In: Experimental Cell Research, Vol. 316, No. 7, 15.04.2010, p. 1271-1281.

Research output: Contribution to journalArticle

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