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  1. Article: Differentiation capacity and maintenance of differentiated phenotypes of human mesenchymal stromal cells cultured on two distinct types of 3D polymeric scaffolds

    Leferink, A. M / Santos, D / Karperien, M / Truckenmüller, R. K / van Blitterswijk, C. A / Moroni, L

    Integrative biology. 2015 Nov. 30, v. 7, no. 12

    2015  

    Abstract: Many studies have shown the influence of soluble factors and material properties on the differentiation capacity of mesenchymal stromal cells (MSCs) cultured as monolayers. These types of two-dimensional (2D) studies can be used as simplified models to ... ...

    Abstract Many studies have shown the influence of soluble factors and material properties on the differentiation capacity of mesenchymal stromal cells (MSCs) cultured as monolayers. These types of two-dimensional (2D) studies can be used as simplified models to understand cell processes related to stem cell sensing and mechano-transduction in a three-dimensional (3D) context. For several other mechanisms such as cell–cell signaling, cell proliferation and cell morphology, it is well-known that cells behave differently on a planar surface compared to cells in 3D environments. In classical tissue engineering approaches, a combination of cells, 3D scaffolds and soluble factors are considered as the key ingredients for the generation of mechanically stable 3D tissue constructs. However, when MSCs are used for tissue engineering strategies, little is known about the maintenance of their differentiation potential in 3D scaffolds after the removal of differentiation soluble factors. In this study, the differentiation potential of human MSCs (hMSCs) into the chondrogenic and osteogenic lineages on two distinct 3D scaffolds, additive manufactured electrospun scaffolds, was assessed and compared to conventional 2D culture. Human MSCs cultured in the presence of soluble factors in 3D showed to differentiate to the same extent as hMSCs cultured as 2D monolayers or as scaffold-free pellets, indicating that the two scaffolds do not play a consistent role in the differentiation process. In the case of phenotypic changes, the achieved differentiated phenotype was not maintained after the removal of soluble factors, suggesting that the plasticity of hMSCs is retained in 3D cell culture systems. This finding can have implications for future tissue engineering approaches in which the validation of hMSC differentiation on 3D scaffolds will not be sufficient to ensure the maintenance of the functionality of the cells in the absence of appropriate differentiation signals.
    Keywords bone formation ; cell culture ; cell proliferation ; cell structures ; humans ; ingredients ; mesenchymal stromal cells ; models ; pellets ; phenotype ; phenotypic plasticity ; polymers ; stem cells ; tissue engineering
    Language English
    Dates of publication 2015-1130
    Size p. 1574-1586.
    Publishing place The Royal Society of Chemistry
    Document type Article
    ZDB-ID 2480063-6
    ISSN 1757-9708 ; 1757-9694
    ISSN (online) 1757-9708
    ISSN 1757-9694
    DOI 10.1039/c5ib00177c
    Database NAL-Catalogue (AGRICOLA)

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  2. Article ; Online: Shape-defined solid micro-objects from poly(d,l-lactic acid) as cell-supportive counterparts in bottom-up tissue engineering.

    Leferink, A M / Tibbe, M P / Bossink, E G B M / de Heus, L E / van Vossen, H / van den Berg, A / Moroni, L / Truckenmüller, R K

    Materials today. Bio

    2019  Volume 4, Page(s) 100025

    Abstract: In bottom-up tissue engineering, small modular units of cells and biomaterials are assembled toward ​larger and more complex ones. In conjunction with a new implementation of this approach, a novel method to fabricate microscale objects from biopolymers ... ...

    Abstract In bottom-up tissue engineering, small modular units of cells and biomaterials are assembled toward ​larger and more complex ones. In conjunction with a new implementation of this approach, a novel method to fabricate microscale objects from biopolymers by thermal imprinting on water-soluble sacrificial layers is presented. By this means, geometrically well-defined objects could be obtained without involving toxic agents in the form of photoinitiators. The micro-objects were used as cell-adhesive substrates and cell spacers in engineered tissues created by cell-guided assembly of the objects. Such constructs can be applied both for
    Language English
    Publishing date 2019-08-20
    Publishing country England
    Document type Journal Article
    ISSN 2590-0064
    ISSN (online) 2590-0064
    DOI 10.1016/j.mtbio.2019.100025
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: 3D high throughput screening and profiling of embryoid bodies in thermoformed microwell plates.

    Vrij, E J / Espinoza, S / Heilig, M / Kolew, A / Schneider, M / van Blitterswijk, C A / Truckenmüller, R K / Rivron, N C

    Lab on a chip

    2016  Volume 16, Issue 4, Page(s) 734–742

    Abstract: 3D organoids using stem cells to study development and disease are now widespread. These models are powerful to mimic in vivo situations but are currently associated with high variability and low throughput. For biomedical research, platforms are thus ... ...

    Abstract 3D organoids using stem cells to study development and disease are now widespread. These models are powerful to mimic in vivo situations but are currently associated with high variability and low throughput. For biomedical research, platforms are thus necessary to increase reproducibility and allow high-throughput screens (HTS). Here, we introduce a microwell platform, integrated in standard culture plates, for functional HTS. Using micro-thermoforming, we form round-bottom microwell arrays from optically clear cyclic olefin polymer films, and assemble them with bottom-less 96-well plates. We show that embryonic stem cells aggregate faster and more reproducibly (centricity, circularity) as compared to a state-of-the-art microwell array. We then run a screen of a chemical library to direct differentiation into primitive endoderm (PrE) and, using on-chip high content imaging (HCI), we identify molecules, including regulators of the cAMP pathway, regulating tissue size, morphology and PrE gene activity. We propose that this platform will benefit to the systematic study of organogenesis in vitro.
    MeSH term(s) Animals ; Cell Aggregation/drug effects ; Cell Line ; Cyclic AMP/metabolism ; Cyclic AMP-Dependent Protein Kinases/metabolism ; Drug Evaluation, Preclinical/methods ; Embryoid Bodies/cytology ; Embryoid Bodies/drug effects ; Enzyme Activation/drug effects ; Gene Expression Regulation, Enzymologic/drug effects ; High-Throughput Screening Assays/methods ; Kinetics ; Mice ; Microtechnology/methods ; Protein Kinase Inhibitors/chemistry ; Protein Kinase Inhibitors/pharmacology ; Receptor, Platelet-Derived Growth Factor alpha/metabolism ; Reproducibility of Results ; Temperature
    Chemical Substances Protein Kinase Inhibitors ; Cyclic AMP (E0399OZS9N) ; Receptor, Platelet-Derived Growth Factor alpha (EC 2.7.10.1) ; Cyclic AMP-Dependent Protein Kinases (EC 2.7.11.11)
    Language English
    Publishing date 2016-02-21
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2056646-3
    ISSN 1473-0189 ; 1473-0197
    ISSN (online) 1473-0189
    ISSN 1473-0197
    DOI 10.1039/c5lc01499a
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Differentiation capacity and maintenance of differentiated phenotypes of human mesenchymal stromal cells cultured on two distinct types of 3D polymeric scaffolds.

    Leferink, A M / Santos, D / Karperien, M / Truckenmüller, R K / van Blitterswijk, C A / Moroni, L

    Integrative biology : quantitative biosciences from nano to macro

    2015  Volume 7, Issue 12, Page(s) 1574–1586

    Abstract: Many studies have shown the influence of soluble factors and material properties on the differentiation capacity of mesenchymal stromal cells (MSCs) cultured as monolayers. These types of two-dimensional (2D) studies can be used as simplified models to ... ...

    Abstract Many studies have shown the influence of soluble factors and material properties on the differentiation capacity of mesenchymal stromal cells (MSCs) cultured as monolayers. These types of two-dimensional (2D) studies can be used as simplified models to understand cell processes related to stem cell sensing and mechano-transduction in a three-dimensional (3D) context. For several other mechanisms such as cell-cell signaling, cell proliferation and cell morphology, it is well-known that cells behave differently on a planar surface compared to cells in 3D environments. In classical tissue engineering approaches, a combination of cells, 3D scaffolds and soluble factors are considered as the key ingredients for the generation of mechanically stable 3D tissue constructs. However, when MSCs are used for tissue engineering strategies, little is known about the maintenance of their differentiation potential in 3D scaffolds after the removal of differentiation soluble factors. In this study, the differentiation potential of human MSCs (hMSCs) into the chondrogenic and osteogenic lineages on two distinct 3D scaffolds, additive manufactured electrospun scaffolds, was assessed and compared to conventional 2D culture. Human MSCs cultured in the presence of soluble factors in 3D showed to differentiate to the same extent as hMSCs cultured as 2D monolayers or as scaffold-free pellets, indicating that the two scaffolds do not play a consistent role in the differentiation process. In the case of phenotypic changes, the achieved differentiated phenotype was not maintained after the removal of soluble factors, suggesting that the plasticity of hMSCs is retained in 3D cell culture systems. This finding can have implications for future tissue engineering approaches in which the validation of hMSC differentiation on 3D scaffolds will not be sufficient to ensure the maintenance of the functionality of the cells in the absence of appropriate differentiation signals.
    MeSH term(s) Alkaline Phosphatase/metabolism ; Cell Culture Techniques ; Cell Dedifferentiation ; Cell Differentiation ; Chondrogenesis ; Extracellular Matrix/metabolism ; Humans ; Mechanotransduction, Cellular ; Mesenchymal Stromal Cells/cytology ; Mesenchymal Stromal Cells/physiology ; Microscopy, Electron, Scanning ; Osteogenesis ; Phenotype ; Polymers/chemistry ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Tissue Engineering ; Tissue Scaffolds/chemistry
    Chemical Substances Polymers ; RNA, Messenger ; Alkaline Phosphatase (EC 3.1.3.1)
    Language English
    Publishing date 2015-12
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2480063-6
    ISSN 1757-9708 ; 1757-9694
    ISSN (online) 1757-9708
    ISSN 1757-9694
    DOI 10.1039/c5ib00177c
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: A modular versatile chip carrier for high-throughput screening of cell-biomaterial interactions.

    Unadkat, H V / Rewagad, R R / Hulsman, M / Hulshof, G F B / Truckenmüller, R K / Stamatialis, D F / Reinders, M J T / Eijkel, J C T / van den Berg, A / van Blitterswijk, C A / de Boer, J

    Journal of the Royal Society, Interface

    2012  Volume 10, Issue 78, Page(s) 20120753

    Abstract: The field of biomaterials research is witnessing a steady rise in high-throughput screening approaches, comprising arrays of materials of different physico-chemical composition in a chip format. Even though the cell arrays provide many benefits in terms ... ...

    Abstract The field of biomaterials research is witnessing a steady rise in high-throughput screening approaches, comprising arrays of materials of different physico-chemical composition in a chip format. Even though the cell arrays provide many benefits in terms of throughput, they also bring new challenges. One of them is the establishment of robust homogeneous cell seeding techniques and strong control over cell culture, especially for long time periods. To meet these demands, seeding cells with low variation per tester area is required, in addition to robust cell culture parameters. In this study, we describe the development of a modular chip carrier which represents an important step in standardizing cell seeding and cell culture conditions in array formats. Our carrier allows flexible and controlled cell seeding and subsequent cell culture using dynamic perfusion. To demonstrate the application of our device, we successfully cultured and evaluated C2C12 premyoblast cell viability under dynamic conditions for a period of 5 days using an automated pipeline for image acquisition and analysis. In addition, using computational fluid dynamics, lactate and BMP-2 as model molecules, we estimated that there is good exchange of nutrients and metabolites with the flowing medium, whereas no cross-talk between adjacent TestUnits should be expected. Moreover, the shear stresses to the cells can be tailored uniformly over the entire chip area. Based on these findings, we believe our chip carrier may be a versatile tool for high-throughput cell experiments in biomaterials sciences.
    MeSH term(s) Biocompatible Materials ; Bone Morphogenetic Protein 2/metabolism ; Cell Culture Techniques ; Cell Line ; Humans ; Lactic Acid/metabolism ; Materials Testing/instrumentation ; Materials Testing/methods ; Microfluidic Analytical Techniques/instrumentation ; Microfluidic Analytical Techniques/methods ; Myoblasts/cytology ; Myoblasts/metabolism ; Stress, Physiological/physiology
    Chemical Substances BMP2 protein, human ; Biocompatible Materials ; Bone Morphogenetic Protein 2 ; Lactic Acid (33X04XA5AT)
    Language English
    Publishing date 2012-11-14
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2156283-0
    ISSN 1742-5662 ; 1742-5689
    ISSN (online) 1742-5662
    ISSN 1742-5689
    DOI 10.1098/rsif.2012.0753
    Database MEDical Literature Analysis and Retrieval System OnLINE

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