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

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-{beta}1, 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,more » along with a contribution to the formation of extracellular matrix.« less

Authors:
 [1];  [2]; ;  [1];  [3];  [4]
  1. Institute for Science and Technology in Medicine, Keele University RJAH Orthopaedic Hospital, Oswestry, SY10 7AG (United Kingdom)
  2. (United Kingdom)
  3. Kroto Research Institute and Centre for Nanoscience and Technology, Sheffield University, Sheffield, S1 2UE (United Kingdom)
  4. School of Life and Health Science, Aston University, Aston Triangle, Birmingham, B4 7EJ (United Kingdom)
Publication Date:
OSTI Identifier:
22212072
Resource Type:
Journal Article
Journal Name:
Experimental Cell Research
Additional Journal Information:
Journal Volume: 316; Journal Issue: 7; Other Information: Copyright (c) 2010 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0014-4827
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; BIOLOGICAL REPAIR; CELL PROLIFERATION; COLLAGEN; FIBROBLASTS; HEALING; IN VITRO; MICROSCOPY; SKIN; STEM CELLS; WOUNDS

Citation Formats

Walter, M.N.M., School of Life and Health Science, Aston University, Aston Triangle, Birmingham, B4 7EJ, Wright, K.T., Fuller, H.R., MacNeil, S., and Johnson, W.E.B., E-mail: w.e.johnson@aston.ac.uk. Mesenchymal stem cell-conditioned medium accelerates skin wound healing: An in vitro study of fibroblast and keratinocyte scratch assays. United States: N. p., 2010. Web. doi:10.1016/J.YEXCR.2010.02.026.
Walter, M.N.M., School of Life and Health Science, Aston University, Aston Triangle, Birmingham, B4 7EJ, Wright, K.T., Fuller, H.R., MacNeil, S., & Johnson, W.E.B., E-mail: w.e.johnson@aston.ac.uk. Mesenchymal stem cell-conditioned medium accelerates skin wound healing: An in vitro study of fibroblast and keratinocyte scratch assays. United States. doi:10.1016/J.YEXCR.2010.02.026.
Walter, M.N.M., School of Life and Health Science, Aston University, Aston Triangle, Birmingham, B4 7EJ, Wright, K.T., Fuller, H.R., MacNeil, S., and Johnson, W.E.B., E-mail: w.e.johnson@aston.ac.uk. Thu . "Mesenchymal stem cell-conditioned medium accelerates skin wound healing: An in vitro study of fibroblast and keratinocyte scratch assays". United States. doi:10.1016/J.YEXCR.2010.02.026.
@article{osti_22212072,
title = {Mesenchymal stem cell-conditioned medium accelerates skin wound healing: An in vitro study of fibroblast and keratinocyte scratch assays},
author = {Walter, M.N.M. and School of Life and Health Science, Aston University, Aston Triangle, Birmingham, B4 7EJ and Wright, K.T. and Fuller, H.R. and MacNeil, S. and Johnson, W.E.B., E-mail: w.e.johnson@aston.ac.uk},
abstractNote = {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-{beta}1, 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.},
doi = {10.1016/J.YEXCR.2010.02.026},
journal = {Experimental Cell Research},
issn = {0014-4827},
number = 7,
volume = 316,
place = {United States},
year = {2010},
month = {4}
}