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  • 1.
    Fromell, Karin
    et al.
    Uppsala University.
    Duhrkop, Claudia
    Uppsala University.
    Johansson, Ulrika
    Linnéuniversitetet, Fakulteten för Hälso- och livsvetenskap (FHL), Institutionen för kemi och biomedicin (KOB).
    Nilsson Ekdahl, Kristina
    Linnéuniversitetet, Fakulteten för Hälso- och livsvetenskap (FHL), Institutionen för kemi och biomedicin (KOB). Uppsala University.
    Nilsson, Bo
    Uppsala University.
    Forms of contact-activated C3 associated with AP convertase formation2017Ingår i: Molecular Immunology, ISSN 0161-5890, E-ISSN 1872-9142, Vol. 89, s. 141-141Artikel i tidskrift (Övrigt vetenskapligt)
  • 2.
    Fromell, Karin
    et al.
    Uppsala University.
    Johansson, Ulrika
    Linnéuniversitetet, Fakulteten för Hälso- och livsvetenskap (FHL), Institutionen för kemi och biomedicin (KOB). Uppsala University.
    Duhrkop, Claudia
    Uppsala University.
    Adler, Anna
    Uppsala University.
    Usterud, Emma
    Uppsala University.
    Hamad, Osama A.
    Uppsala University.
    Nilsson Ekdahl, Kristina
    Linnéuniversitetet, Fakulteten för Hälso- och livsvetenskap (FHL), Institutionen för kemi och biomedicin (KOB). Uppsala University.
    Nilsson, Bo
    Uppsala University.
    Generation of an alternative pathway convertase by contact-activated C3 is dependent on the conformation of C32018Ingår i: Molecular Immunology, ISSN 0161-5890, E-ISSN 1872-9142, Vol. 102, s. 193-193Artikel i tidskrift (Övrigt vetenskapligt)
  • 3.
    Johansson, Ulrika
    et al.
    Swedish University of Agricultural Sciences ; Karolinska Institutet.
    Ria, Massimiliano
    Karolinska Institutet.
    Åvall, Karin
    Karolinska Institutet.
    Shalaly, Nancy Dekki
    KTH Royal Institute of Technology.
    Zaisev, Sergei V.
    Karolinska Institutet.
    Berggren, Per-Olof
    Karolinska Institutet.
    Hedhammar, My
    Karolinska Institutet.
    Pancreatic Islet Survival and Engraftment Is Promoted by Culture on Functionalized Spider Silk Matrices2015Ingår i: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 19, nr 10, s. 1-21Artikel i tidskrift (Refereegranskat)
    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.

  • 4.
    Johansson, Ulrika
    et al.
    Linnéuniversitetet, Fakulteten för Hälso- och livsvetenskap (FHL), Institutionen för kemi och biomedicin (KOB). KTH Royal Instute of Technology, Sweden;Uppsala University, Sweden.
    Widhe, Mona
    KTH Royal Instute of Technology, Sweden.
    Shalaly, Nancy Dekki
    KTH Royal Instute of Technology, Sweden.
    Arregui, Irene Linares
    KTH Royal Instute of Technology, Sweden.
    Nileback, Linnea
    KTH Royal Instute of Technology, Sweden.
    Tasiopoulos, Christos Panagiotis
    KTH Royal Instute of Technology, Sweden.
    Åstrand, Carolina
    KTH Royal Instute of Technology, Sweden.
    Berggren, Per-Olof
    Karolinska Institutet, Sweden;Karolinska University Hospital, Sweden.
    Gasser, Christian
    KTH Royal Instute of Technology, Sweden.
    Hedhammar, My
    KTH Royal Instute of Technology, Sweden.
    Assembly of functionalized silk together with cells to obtain proliferative 3D cultures integrated in a network of ECM-like microfibers2019Ingår i: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 9, s. 1-13, artikel-id 6291Artikel i tidskrift (Refereegranskat)
    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.

  • 5.
    Shalaly, Nancy Dekki
    et al.
    KTH Royal Institute of Technology.
    Ria, Massimiliano
    Karolinska Institutet.
    Johansson, Ulrika
    KTH Royal Institute of Technology.
    Åvall, Karin
    Karolinska Institutet.
    Berggren, Per-Olof
    Karolinska Institutet.
    Hedhammar, My
    KTH Royal Institute of Technology ; Swedish University of Agricultural Sciences.
    Silk matrices promote formation of insulin-secreting islet-like clusters2016Ingår i: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 90, s. 50-61Artikel i tidskrift (Refereegranskat)
    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.

  • 6.
    Widhe, Mona
    et al.
    Swedish University of Agricultural Sciences.
    Johansson, Ulrika
    Swedish University of Agricultural Sciences.
    Hillerdahl, Carl-Olof
    Swedish University of Agricultural Sciences.
    Hedhammar, My
    Swedish University of Agricultural Sciences.
    Recombinant spider silk with cell binding motifs for specific adherence of cells2013Ingår i: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 34, nr 33, s. 8223-8234Artikel i tidskrift (Refereegranskat)
    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.

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