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Johansson, Ulrika
Publications (6 of 6) Show all publications
Johansson, U., Widhe, M., Shalaly, N. D., Arregui, I. L., Nileback, L., Tasiopoulos, C. P., . . . Hedhammar, M. (2019). Assembly of functionalized silk together with cells to obtain proliferative 3D cultures integrated in a network of ECM-like microfibers. Scientific Reports, 9, 1-13, Article ID 6291.
Open this publication in new window or tab >>Assembly of functionalized silk together with cells to obtain proliferative 3D cultures integrated in a network of ECM-like microfibers
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2019 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 9, p. 1-13, article id 6291Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
Nature Publishing Group, 2019
National Category
Cell Biology
Research subject
Natural Science, Cell and Organism Biology
Identifiers
urn:nbn:se:lnu:diva-82537 (URN)10.1038/s41598-019-42541-y (DOI)000465001600045 ()31000733 (PubMedID)
Available from: 2019-05-14 Created: 2019-05-14 Last updated: 2019-05-14Bibliographically approved
Fromell, K., Johansson, U., Duhrkop, C., Adler, A., Usterud, E., Hamad, O. A., . . . Nilsson, B. (2018). Generation of an alternative pathway convertase by contact-activated C3 is dependent on the conformation of C3. Paper presented at 27th International Complement Workshop (ICW), SEP 16-20, 2018, Santa Fe, NM. Molecular Immunology, 102, 193-193
Open this publication in new window or tab >>Generation of an alternative pathway convertase by contact-activated C3 is dependent on the conformation of C3
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2018 (English)In: Molecular Immunology, ISSN 0161-5890, E-ISSN 1872-9142, Vol. 102, p. 193-193Article in journal, Meeting abstract (Other academic) Published
Place, publisher, year, edition, pages
Elsevier, 2018
Keywords
Complement C3
National Category
Immunology
Research subject
Biomedical Sciences, Immunology
Identifiers
urn:nbn:se:lnu:diva-78410 (URN)10.1016/j.molimm.2018.06.167 (DOI)000445313600163 ()
Conference
27th International Complement Workshop (ICW), SEP 16-20, 2018, Santa Fe, NM
Available from: 2018-10-22 Created: 2018-10-22 Last updated: 2018-11-02Bibliographically approved
Fromell, K., Duhrkop, C., Johansson, U., Nilsson Ekdahl, K. & Nilsson, B. (2017). Forms of contact-activated C3 associated with AP convertase formation. Paper presented at 16th European Meeting on Complement in Human Disease (EMCHD), SEP 08-12, 2017, Copenhagen, DENMARK. Molecular Immunology, 89, 141-141
Open this publication in new window or tab >>Forms of contact-activated C3 associated with AP convertase formation
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2017 (English)In: Molecular Immunology, ISSN 0161-5890, E-ISSN 1872-9142, Vol. 89, p. 141-141Article in journal, Meeting abstract (Other academic) Published
National Category
Immunology
Research subject
Biomedical Sciences, Immunology
Identifiers
urn:nbn:se:lnu:diva-68147 (URN)10.1016/j.molimm.2017.06.085 (DOI)000410014500065 ()
Conference
16th European Meeting on Complement in Human Disease (EMCHD), SEP 08-12, 2017, Copenhagen, DENMARK
Available from: 2017-10-02 Created: 2017-10-02 Last updated: 2018-11-16Bibliographically approved
Shalaly, N. D., Ria, M., Johansson, U., Åvall, K., Berggren, P.-O. & Hedhammar, M. (2016). Silk matrices promote formation of insulin-secreting islet-like clusters. Biomaterials, 90, 50-61
Open this publication in new window or tab >>Silk matrices promote formation of insulin-secreting islet-like clusters
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2016 (English)In: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 90, p. 50-61Article in journal (Refereed) Published
Abstract [en]

Ex vivo expansion of endocrine cells constitutes an interesting alternative to be able to match the unmet need of transplantable pancreatic islets. However, endocrine cells become fragile once removed from their extracellular matrix (ECM) and typically become senescent and loose insulin expression during conventional 2D culture. Herein we develop a protocol where 3D silk matrices functionalized with ECM-derived motifs are used for generation of insulin-secreting islet-like clusters from mouse and human primary cells. The obtained clusters were shown to attain an islet-like spheroid shape and to maintain functional insulin release upon glucose stimulation in vitro. Furthermore, in vivo imaging of transplanted murine clusters showed engraftment with increasing vessel formation during time. There was no sign of cell death and the clusters maintained or increased in size throughout the period, thus suggesting a suitable cluster size for transplantation.

Keywords
Recombinant spider silk, Pancreatic islets, Islet-like clusters, Insulin release, In vivo imaging, Vascularization
National Category
Cell and Molecular Biology
Research subject
Natural Science
Identifiers
urn:nbn:se:lnu:diva-61320 (URN)10.1016/j.biomaterials.2016.03.006 (DOI)
Available from: 2017-03-13 Created: 2017-03-13 Last updated: 2018-01-13Bibliographically approved
Johansson, U., Ria, M., Åvall, K., Shalaly, N. D., Zaisev, S. V., Berggren, P.-O. & Hedhammar, M. (2015). Pancreatic Islet Survival and Engraftment Is Promoted by Culture on Functionalized Spider Silk Matrices. PLoS ONE, 19(10), 1-21
Open this publication in new window or tab >>Pancreatic Islet Survival and Engraftment Is Promoted by Culture on Functionalized Spider Silk Matrices
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2015 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 19, no 10, p. 1-21Article in journal (Refereed) Published
Abstract [en]

Transplantation of pancreatic islets is one approach for treatment of diabetes, however, hampered by the low availability of viable islets. Islet isolation leads to disruption of the environment surrounding the endocrine cells, which contributes to eventual cell death. The reestablishment of this environment is vital, why we herein investigated the possibility of using recombinant spider silk to support islets in vitro after isolation. The spider silk protein 4RepCT was formulated into three different formats; 2D-film, fiber mesh and 3D-foam, in order to provide a matrix that can give the islets physical support in vitro. Moreover, cell-binding motifs from laminin were incorporated into the silk protein in order to create matrices that mimic the natural cell environment. Pancreatic mouse islets were thoroughly analyzed for adherence, necrosis and function after in vitro maintenance on the silk matrices. To investigate their suitability for transplantation, we utilized an eye model which allows in vivo imaging of engraftment. Interestingly, islets that had been maintained on silk foam during in vitro culture showed improved revascularization. This coincided with the observation of preserved islet architecture with endothelial cells present after in vitro culture on silk foam. Selected matrices were further evaluated for long-term preservation of human islets. Matrices with the cell-binding motif RGD improved human islet maintenance (from 36% to 79%) with preserved islets architecture and function for over 3 months in vitro. The islets established cell-matrix contacts and formed vessel-like structures along the silk. Moreover, RGD matrices promoted formation of new, insulin-positive islet-like clusters that were connected to the original islets via endothelial cells. On silk matrices with islets from younger donors (<35 year), the amount of newly formed islet-like clusters found after 1 month in culture were almost double compared to the initial number of islets added.

National Category
Cell and Molecular Biology
Research subject
Natural Science
Identifiers
urn:nbn:se:lnu:diva-61319 (URN)10.1371/journal.pone.0130169 (DOI)
Available from: 2017-03-13 Created: 2017-03-13 Last updated: 2018-01-13Bibliographically approved
Widhe, M., Johansson, U., Hillerdahl, C.-O. & Hedhammar, M. (2013). Recombinant spider silk with cell binding motifs for specific adherence of cells. Biomaterials, 34(33), 8223-8234
Open this publication in new window or tab >>Recombinant spider silk with cell binding motifs for specific adherence of cells
2013 (English)In: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 34, no 33, p. 8223-8234Article in journal (Refereed) Published
Abstract [en]

Silk matrices have previously been shown to possess general properties governing cell viability. However, many cell types also require specific adhesion sites for successful in vitro culture. Herein, we have shown that cell binding motifs can be genetically fused to a partial spider silk protein, 4RepCT, without affecting its ability to self-assemble into stable matrices directly in a physiological-like buffer. The incorporated motifs were exposed in the formed matrices, and available for binding of integrins. Four different human primary cell types; fibroblasts, keratinocytes, endothelial cells and Schwann cells, were applied to the matrices and investigated under serum-free culture conditions. Silk matrices with cell binding motifs, especially RGD, were shown to promote early adherence of cells, which formed stress fibers and distinct focal adhesion points. Schwann cells acquired most spread-out morphology on silk matrices with IKVAV, where significantly more viable cells were found, also when compared to wells coated with laminin. This strategy is thus suitable for development of matrices that allow screening of various cell binding motifs and their effect on different cell types.

Keywords
Spider silk, Cell culture matrices, Cell adhesion, Integrin
National Category
Cell and Molecular Biology
Research subject
Natural Science
Identifiers
urn:nbn:se:lnu:diva-61318 (URN)10.1016/j.biomaterials.2013.07.058 (DOI)
Available from: 2017-03-13 Created: 2017-03-13 Last updated: 2018-01-13Bibliographically approved
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