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Title: Extracellular matrix components direct porcine muscle stem cell behavior

Abstract

In muscle tissue, extracellular matrix proteins, together with the vasculature system, muscle-residence cells and muscle fibers, create the niche for muscle stem cells. The niche is important in controlling proliferation and directing differentiation of muscle stem cells to sustain muscle tissue. Mimicking the extracellular muscle environment improves tools exploring the behavior of primary muscle cells. Optimizing cell culture conditions to maintain muscle commitment is important in stem cell-based studies concerning toxicology screening, ex vivo skeletal muscle tissue engineering and in the enhancement of clinical efficiency. We used the muscle extracellular matrix proteins collagen type I, fibronectin, laminin, and also gelatin and Matrigel as surface coatings of tissue culture plastic to resemble the muscle extracellular matrix. Several important factors that determine myogenic commitment of the primary muscle cells were characterized by quantitative real-time RT-PCR and immunofluorescence. Adhesion of high PAX7 expressing satellite cells was improved if the cells were cultured on fibronectin or laminin coatings. Cells cultured on Matrigel and laminin coatings showed dominant integrin expression levels and exhibited an activated Wnt pathway. Under these conditions both stem cell proliferation and myogenic differentiation capacity were superior if compared to cells cultured on collagen type I, fibronectin and gelatin. In conclusion, Matrigelmore » and laminin are the preferred coatings to sustain the proliferation and myogenic differentiation capacity of the primary porcine muscle stem cells, when cells are removed from their natural environment for in vitro culture.« less

Authors:
 [1];  [2];  [1]
  1. Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, 3584 CM, Utrecht (Netherlands)
  2. Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL, Utrecht (Netherlands)
Publication Date:
OSTI Identifier:
22209853
Resource Type:
Journal Article
Journal Name:
Experimental Cell Research
Additional Journal Information:
Journal Volume: 316; Journal Issue: 3; Other Information: Copyright (c) 2009 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; ANIMAL TISSUES; CELL CULTURES; COLLAGEN; GELATIN; MUSCLES; MYOSIN; OPTIMIZATION; PHOSPHATES; POLYMERASE CHAIN REACTION; STEM CELLS; SWINE

Citation Formats

Wilschut, Karlijn J., Haagsman, Henk P., and Roelen, Bernard A.J., E-mail: b.a.j.roelen@uu.nl. Extracellular matrix components direct porcine muscle stem cell behavior. United States: N. p., 2010. Web. doi:10.1016/J.YEXCR.2009.10.014.
Wilschut, Karlijn J., Haagsman, Henk P., & Roelen, Bernard A.J., E-mail: b.a.j.roelen@uu.nl. Extracellular matrix components direct porcine muscle stem cell behavior. United States. doi:10.1016/J.YEXCR.2009.10.014.
Wilschut, Karlijn J., Haagsman, Henk P., and Roelen, Bernard A.J., E-mail: b.a.j.roelen@uu.nl. Mon . "Extracellular matrix components direct porcine muscle stem cell behavior". United States. doi:10.1016/J.YEXCR.2009.10.014.
@article{osti_22209853,
title = {Extracellular matrix components direct porcine muscle stem cell behavior},
author = {Wilschut, Karlijn J. and Haagsman, Henk P. and Roelen, Bernard A.J., E-mail: b.a.j.roelen@uu.nl},
abstractNote = {In muscle tissue, extracellular matrix proteins, together with the vasculature system, muscle-residence cells and muscle fibers, create the niche for muscle stem cells. The niche is important in controlling proliferation and directing differentiation of muscle stem cells to sustain muscle tissue. Mimicking the extracellular muscle environment improves tools exploring the behavior of primary muscle cells. Optimizing cell culture conditions to maintain muscle commitment is important in stem cell-based studies concerning toxicology screening, ex vivo skeletal muscle tissue engineering and in the enhancement of clinical efficiency. We used the muscle extracellular matrix proteins collagen type I, fibronectin, laminin, and also gelatin and Matrigel as surface coatings of tissue culture plastic to resemble the muscle extracellular matrix. Several important factors that determine myogenic commitment of the primary muscle cells were characterized by quantitative real-time RT-PCR and immunofluorescence. Adhesion of high PAX7 expressing satellite cells was improved if the cells were cultured on fibronectin or laminin coatings. Cells cultured on Matrigel and laminin coatings showed dominant integrin expression levels and exhibited an activated Wnt pathway. Under these conditions both stem cell proliferation and myogenic differentiation capacity were superior if compared to cells cultured on collagen type I, fibronectin and gelatin. In conclusion, Matrigel and laminin are the preferred coatings to sustain the proliferation and myogenic differentiation capacity of the primary porcine muscle stem cells, when cells are removed from their natural environment for in vitro culture.},
doi = {10.1016/J.YEXCR.2009.10.014},
journal = {Experimental Cell Research},
issn = {0014-4827},
number = 3,
volume = 316,
place = {United States},
year = {2010},
month = {2}
}