Assembly of functionalized silk together with cells to obtain proliferative 3D cultures integrated in a network of ECM-like microfibers.

Johansson, Ulrika; Widhe, Mona; Shalaly, Nancy Dekki; Arregui, Irene Linares; Nilebäck, Linnea; Tasiopoulos, Christos Panagiotis; Åstrand, Carolina; Berggren, Per-Olof; Gasser, Christian; Hedhammar, My

Johansson, Ulrika; Widhe, Mona; Shalaly, Nancy Dekki; Arregui, Irene Linares; Nilebäck, Linnea; Tasiopoulos, Christos Panagiotis; Åstrand, Carolina; Berggren, Per-Olof; Gasser, Christian; Hedhammar, My.

Afiliación

Johansson U; Division of Protein Technology, School of Biotechnology, KTH Royal Institute of Technology, SE-106 91, Stockholm, Sweden.
Widhe M; Linnæus Center of Biomaterials Chemistry, Linnæus University, Kalmar, Sweden.
Shalaly ND; Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
Arregui IL; Division of Protein Technology, School of Biotechnology, KTH Royal Institute of Technology, SE-106 91, Stockholm, Sweden.
Nilebäck L; Division of Protein Technology, School of Biotechnology, KTH Royal Institute of Technology, SE-106 91, Stockholm, Sweden.
Tasiopoulos CP; Department of Solid Mechanics, KTH Royal Institute of Technology, SE-106 91, Stockholm, Sweden.
Åstrand C; Division of Protein Technology, School of Biotechnology, KTH Royal Institute of Technology, SE-106 91, Stockholm, Sweden.
Berggren PO; Division of Protein Technology, School of Biotechnology, KTH Royal Institute of Technology, SE-106 91, Stockholm, Sweden.
Gasser C; Division of Protein Technology, School of Biotechnology, KTH Royal Institute of Technology, SE-106 91, Stockholm, Sweden.
Hedhammar M; The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital, S-171 76, Stockholm, Sweden.

Sci Rep ; 9(1): 6291, 2019 04 18.

Article en En | MEDLINE | ID: mdl-31000733

RESUMEN

Tissues are built of cells integrated in an extracellular matrix (ECM) which provides a three-dimensional (3D) microfiber 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 networks of microfibers 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 microfibers. In the resulting 3D scaffold, the cells are highly proliferative and spread out more efficiently than when encapsulated in a hydrogel. 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 microfibers formed in a bundle with integrated cells are both strong and extendable, with mechanical properties similar to that of artery walls. The described method enables differentiation of stem cells in 3D as well as 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 ECM-like network, with potential as base for engineering of functional tissue.

Asunto(s)

Matriz Extracelular/genética; Fibronectinas/genética; Proteínas Recombinantes/genética; Seda/genética; Animales; Adhesión Celular/genética; Técnicas de Cultivo de Célula; Diferenciación Celular/genética; Proliferación Celular/genética; Matriz Extracelular/ultraestructura; Fibronectinas/química; Fibronectinas/ultraestructura; Ingeniería Genética; Humanos; Hidrogeles/química; Proteínas Recombinantes/ultraestructura; Seda/ultraestructura; Células Madre/metabolismo

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Recombinantes / Fibronectinas / Seda / Matriz Extracelular Límite: Animals / Humans Idioma: En Revista: Sci Rep Año: 2019 Tipo del documento: Article País de afiliación: Suecia Pais de publicación: Reino Unido

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