Improved proliferation and differentiation capacity of human mesenchymal stromal cells cultured with basement-membrane extracellular matrix proteins |
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| Authors: | Ulrich Lindner Jan Kramer Jochen Behrends Birgit Driller Nils-Ole Wendler Florian Boehrnsen Jürgen Rohwedel Peter Schlenke |
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| Institution: | 1. Medical Department I, Division of Nephrology and Transplantation Unit and;2. Department of Virology and Cell Biology, University of Luebeck, Luebeck, Germany;3. Junior Research Group Molecular Infection Biology, Leibniz Research Center Borstel, Borstel, Germany;4. Institute of Immunology and Transfusion Medicine;5. Department of Orthopaedic Surgery, University of Luebeck, Luebeck, Germany;6. Institute of Transfusion Medicine und Transplantation Immunology, Westphalian Wilhelms-University Muenster, Muenster, Germany;1. Key Laboratory of Modern Acoustics, Ministry of Education, Nanjing University, Nanjing 210093, China;2. Institute of Traumatology and Orthopedics, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, China;3. Department of Electronic Information, Nantong University, Nantong 226019, China;4. Department of Physics, University of Vermont, Burlington, VT 05405, USA;5. The State Key Laboratory of Acoustics, Chinese Academy of Science, Beijing 10080, China;3. Department of Mechanical Engineering, University of New Mexico, Albuquerque, NM 87131, USA |
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| Abstract: | Background aimsIn vitro cultured mesenchymal stromal cells (MSC) are characterized by a short proliferative lifespan, an increasing loss of proliferation capacity and progressive reduction of differentiation potential. Laminin-1, laminin-5, collagen IV and fibronectin are important constituents of the basement membrane extracellular matrix (ECM) that are involved in a variety of cellular activities, including cell attachment and motility.Methods and resultsThe in vitro proliferation capacity of MSC was significantly improved when the cells were incubated in the presence of basement membrane ECM proteins. For example, a mixture of proteins improved proliferation capacity 250-fold in comparison with standard conditions after five passages. Furthermore, in colony-forming unit–fibroblast (CFU-F) assays colony numbers and size were significantly extended. Blocking specific integrin cell-surface receptors, positive effects on the proliferation capacity of MSC were inhibited. Additionally, when MSC were co-cultivated with ECM proteins, cells maintained their multipotential differentiation capacity throughout many culture passages in comparison with cells cultivated on plastic. However, expansion of MSC on laminin-5 suppressed any subsequent chondrogenic differentiation.ConclusionsOur results suggest that expansion of bone marrow-derived MSC in the presence of ECM proteins is a powerful approach for generating large numbers of MSC, showing a prolonged capacity to differentiate into mesodermal cell lineages, with the exception of the lack of chondrogenesis by using laminin-5 coating. |
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