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Silk assembly integrates cells into a 3D fibrillar network that promotes cell spreading and proliferation

Ulrika Johansson, Mona Widhe, Nancy Dekki Shalaly, Irene Linares Arregui, Linnea Nileback, Christos Panagiotis Tasiopoulos, Carolina Astrand, Per-Olof Berggren, Christian Gasser, My Hedhammar
doi: https://doi.org/10.1101/403345
Ulrika Johansson
Royal Institute of Technology;
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Mona Widhe
Royal Institute of Technology;
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Nancy Dekki Shalaly
Royal Institute of Technology;
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Irene Linares Arregui
Royal Institute of Technology;
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Linnea Nileback
Royal Institute of Technology;
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Christos Panagiotis Tasiopoulos
Royal Institute of Technology;
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Carolina Astrand
Royal Institute of Technology;
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Per-Olof Berggren
Karolinska Institutet
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Christian Gasser
Royal Institute of Technology;
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My Hedhammar
Royal Institute of Technology;
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  • For correspondence: myh@kth.se
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Abstract

Tissues are built of cells integrated in an extracellular matrix (ECM) which provides a three-dimensional (3D) fibrillar network with specific sites for cell anchorage. By genetic engineering, motifs from the ECM can be functionally fused to recombinant silk proteins. Such a silk protein, FN-silk, which harbours a motif from fibronectin, has the ability to self-assemble into fibrillar networks under physiological-like conditions. Herein we describe a method by which mammalian cells are added to the silk solution before assembly, and thereby get uniformly integrated between the formed fibrils. In the resulting 3D scaffold, the cells proliferate and spread out with tissue-like morphology. Elongated cells containing filamentous actin and defined focal adhesion points confirm proper cell attachment to the FN-silk. The cells remain viable in culture for at least 90 days. The method is also scalable to macro-sized 3D cultures. Silk fibers with integrated cells are both strong and extendable, with mechanical properties similar to that of artery walls. The described method enables both differentiation of stem- or precursor cells in 3D and facile co-culture of several different cell types. We show that inclusion of endothelial cells leads to the formation of vessel-like structures throughout the tissue constructs. Hence, silk-assembly in presence of cells constitutes a viable option for 3D culture of cells integrated in a fibrillary ECM-like network, with potential as base for engineering of functional tissue.

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Posted August 29, 2018.
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Silk assembly integrates cells into a 3D fibrillar network that promotes cell spreading and proliferation
Ulrika Johansson, Mona Widhe, Nancy Dekki Shalaly, Irene Linares Arregui, Linnea Nileback, Christos Panagiotis Tasiopoulos, Carolina Astrand, Per-Olof Berggren, Christian Gasser, My Hedhammar
bioRxiv 403345; doi: https://doi.org/10.1101/403345
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Silk assembly integrates cells into a 3D fibrillar network that promotes cell spreading and proliferation
Ulrika Johansson, Mona Widhe, Nancy Dekki Shalaly, Irene Linares Arregui, Linnea Nileback, Christos Panagiotis Tasiopoulos, Carolina Astrand, Per-Olof Berggren, Christian Gasser, My Hedhammar
bioRxiv 403345; doi: https://doi.org/10.1101/403345

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