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Open accessJournal ArticleDOI: 10.1038/S41598-021-84384-6

3D bioprinting of hepatocytes: core-shell structured co-cultures with fibroblasts for enhanced functionality

04 Mar 2021-Scientific Reports (Nature Publishing Group)-Vol. 11, Iss: 1, pp 5130-5130
Abstract: With the aim of understanding and recapitulating cellular interactions of hepatocytes in their physiological microenvironment and to generate an artificial 3D in vitro model, a co-culture system using 3D extrusion bioprinting was developed. A bioink based on alginate and methylcellulose (algMC) was first shown to be suitable for bioprinting of hepatocytes; the addition of Matrigel to algMC enhanced proliferation and morphology of them in monophasic scaffolds. Towards a more complex system that allows studying cellular interactions, we applied core–shell bioprinting to establish tailored 3D co-culture models for hepatocytes. The bioinks were specifically functionalized with natural matrix components (based on human plasma, fibrin or Matrigel) and used to co-print fibroblasts and hepatocytes in a spatially defined, coaxial manner. Fibroblasts acted as supportive cells for co-cultured hepatocytes, stimulating the expression of certain biomarkers of hepatocytes like albumin. Furthermore, matrix functionalization positively influenced both cell types in their respective compartments by enhancing their adhesion, viability, proliferation and function. In conclusion, we established a functional co-culture model with independently tunable compartments for different cell types via core–shell bioprinting. This provides the basis for more complex in vitro models allowing co-cultivation of hepatocytes with other liver-specific cell types to closely resemble the liver microenvironment.

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Topics: 3D bioprinting (67%), Matrigel (51%)
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9 results found


Journal ArticleDOI: 10.1039/D1TB00172H
Baosen Tan1, Shaolei Gan, Xiumei Wang2, Wenyong Liu1  +1 moreInstitutions (2)
Abstract: Over the past decade, 3D bioprinting technology has progressed tremendously in the field of tissue engineering in its ability to fabricate individualized biological constructs with precise geometric designability, which offers us the capability to bridge the divergence between engineered tissue constructs and natural tissues. In this work, we first review the current widely used 3D bioprinting approaches, cells, and materials. Next, the updated applications of this technique in tissue engineering, including bone tissue, cartilage tissue, vascular grafts, skin, neural tissue, heart tissue, liver tissue and lung tissue, are briefly introduced. Then, the prominent advantages of 3D bioprinting in tissue engineering are summarized in detail: rapidly prototyping the customized structure, delivering cell-laden materials with high precision in space, and engineering with a highly controllable microenvironment. The current technical deficiencies of 3D bioprinted constructs in terms of mechanical properties and cell behaviors are afterward illustrated, as well as corresponding improvements. Finally, we conclude with future perspectives about 3D bioprinting in tissue engineering.

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Topics: 3D bioprinting (75%)

4 Citations


Open accessJournal ArticleDOI: 10.1016/J.STEM.2021.07.002
04 Nov 2021-Cell Stem Cell
Abstract: Summary In the liver, ductal cells rarely proliferate during homeostasis but do so transiently after tissue injury. These cells can be expanded as organoids that recapitulate several of the cell-autonomous mechanisms of regeneration but lack the stromal interactions of the native tissue. Here, using organoid co-cultures that recapitulate the ductal-to-mesenchymal cell architecture of the portal tract, we demonstrate that a subpopulation of mouse periportal mesenchymal cells exerts dual control on proliferation of the epithelium. Ductal cell proliferation is either induced and sustained or, conversely, completely abolished, depending on the number of direct mesenchymal cell contacts, through a mechanism mediated, at least in part, by Notch signaling. Our findings expand the concept of the cellular niche in epithelial tissues, whereby not only soluble factors but also cell-cell contacts are the key regulatory cues involved in the control of cellular behaviors, suggesting a critical role for cell-cell contacts during regeneration.

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Topics: Liver cell (59%), Ductal cells (56%), Mesenchymal stem cell (54%) ... read more

2 Citations


Journal ArticleDOI: 10.1016/J.BPRINT.2021.E00158
Maureen T. Ross1, David Kilian2, Anja Lode2, Jiongyu Ren1  +3 moreInstitutions (2)
01 Aug 2021-Bioprinting
Abstract: Moulded hydrogels reinforced with melt-electrowritten (MEW) microfibres to tailor mechanical properties show promise in the articular cartilage regeneration field. We aim to transfer this knowledge to bioprinted constructs of volumetric dimensions towards real anatomical shapes, using a previously established alginate methylcellulose (algMC) blend. We show the potential of algMC to be a bioink for MEW fibre-reinforced 3D printed constructs that have tailorable mechanical properties and support long-term culture of bioprinted human chondrocytes. It is hoped that these composite algMC-PCL scaffolds have potential for use in auricular (ear) cartilage regeneration as an alternative to current reconstructive treatments using autografted rib cartilage or high-density polyethylene implants. Scaffolds of different designs were assessed to determine the effect of the hybrid structure on compressive strength, as well as an initial in vitro experiment with human chondrocytes to assess cell viability, DNA, sGAG, and collagen II content. The inclusion of MEW PCL scaffolds showed up to a 7.5-fold increase in the maximum compressive stress after 7 days of incubation in DMEM compared to algMC only scaffolds. In vitro work showed there was no significant differences in the cell viability between groups after 21 days of culture. However, DNA and sGAG content (relative to cell number) showed an increase in algMC scaffolds reinforced with MEW PCL sheets with 750 μm pores suggesting a more favourable stiffness for chondrocyte extracellular matrix (ECM) deposition. Future work will investigate how the scaffold stiffness changes over time as materials degrade and ECM is deposited as well as further in depth analysis of the biological performance with relevance to auricular cartilage. This microfibre reinforced algMC scaffold provides a promising way to tailor the mechanical properties of bioprinted structures for chondrocyte delivery at clinically relevant dimensions.

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Topics: Chondrocyte (52%)

2 Citations


Open accessJournal ArticleDOI: 10.3390/CANCERS13153651
21 Jul 2021-Cancers
Abstract: Hepatocellular carcinoma (HCC), the most frequent form of primary liver carcinoma, is a heterogenous and complex tumor type with increased incidence, poor prognosis, and high mortality. The actual therapeutic arsenal is narrow and poorly effective, rendering this disease a global health concern. Although considerable progress has been made in terms of understanding the pathogenesis, molecular mechanisms, genetics, and therapeutical approaches, several facets of human HCC remain undiscovered. A valuable and prompt approach to acquire further knowledge about the unrevealed aspects of HCC and novel therapeutic candidates is represented by the application of experimental models. Experimental models (in vivo and in vitro 2D and 3D models) are considered reliable tools to gather data for clinical usability. This review offers an overview of the currently available preclinical models frequently applied for the study of hepatocellular carcinoma in terms of initiation, development, and progression, as well as for the discovery of efficient treatments, highlighting the advantages and the limitations of each model. Furthermore, we also focus on the role played by computational studies (in silico models and artificial intelligence-based prediction models) as promising novel tools in liver cancer research.

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1 Citations


Open accessJournal ArticleDOI: 10.3390/IJMS221910214
Sarah Kammerer1Institutions (1)
Abstract: Drug-induced liver injury (DILI) is the major reason for failures in drug development and withdrawal of approved drugs from the market. Two-dimensional cultures of hepatocytes often fail to reliably predict DILI: hepatoma cell lines such as HepG2 do not reflect important primary-like hepatic properties and primary human hepatocytes (pHHs) dedifferentiate quickly in vitro and are, therefore, not suitable for long-term toxicity studies. More predictive liver in vitro models are urgently required in drug development and compound safety evaluation. This review discusses available human hepatic cell types for in vitro toxicology analysis and their usage in established and emerging three-dimensional (3D) culture systems. Generally, 3D cultures maintain or improve primary hepatic functions (including expression of drug-metabolizing enzymes) of different liver cells for several weeks of culture, thus allowing long-term and repeated-dose toxicity studies. Spheroid cultures of pHHs have been comprehensively tested, but also other cell types such as HepaRG benefit from 3D culture systems. Emerging 3D culture techniques include usage of induced pluripotent stem-cell-derived hepatocytes and primary-like upcyte cells, as well as advanced culture techniques such as microfluidic liver-on-a-chip models. In-depth characterization of existing and emerging 3D hepatocyte technologies is indispensable for successful implementation of such systems in toxicological analysis.

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Topics: In vitro toxicology (52%)

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61 results found


Open accessJournal ArticleDOI: 10.1038/S41598-016-0028-X
Michiru Nishita1, Seung-Yeol Park2, Tadashi Nishio1, Koki Kamizaki1  +8 moreInstitutions (5)
26 Jan 2017-Scientific Reports
Abstract: Signaling through the Ror2 receptor tyrosine kinase promotes invadopodia formation for tumor invasion. Here, we identify intraflagellar transport 20 (IFT20) as a new target of this signaling in tumors that lack primary cilia, and find that IFT20 mediates the ability of Ror2 signaling to induce the invasiveness of these tumors. We also find that IFT20 regulates the nucleation of Golgi-derived microtubules by affecting the GM130-AKAP450 complex, which promotes Golgi ribbon formation in achieving polarized secretion for cell migration and invasion. Furthermore, IFT20 promotes the efficiency of transport through the Golgi complex. These findings shed new insights into how Ror2 signaling promotes tumor invasiveness, and also advance the understanding of how Golgi structure and transport can be regulated.

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Topics: Golgi apparatus (58%), Invadopodia (56%), Intraflagellar transport (51%) ... read more

8,752 Citations


Journal ArticleDOI: 10.1002/ADMA.201302042
Jos Malda1, Jos Malda2, Jetze Visser2, Ferry P.W. Melchels2  +7 moreInstitutions (4)
01 Sep 2013-Advanced Materials
Abstract: With advances in tissue engineering, the possibility of regenerating injured tissue or failing organs has become a realistic prospect for the first time in medical history. Tissue engineering - the combination of bioactive materials with cells to generate engineered constructs that functionally replace lost and/or damaged tissue - is a major strategy to achieve this goal. One facet of tissue engineering is biofabrication, where three-dimensional tissue-like structures composed of biomaterials and cells in a single manufacturing procedure are generated. Cell-laden hydrogels are commonly used in biofabrication and are termed "bioinks". Hydrogels are particularly attractive for biofabrication as they recapitulate several features of the natural extracellular matrix and allow cell encapsulation in a highly hydrated mechanically supportive three-dimensional environment. Additionally, they allow for efficient and homogeneous cell seeding, can provide biologically-relevant chemical and physical signals, and can be formed in various shapes and biomechanical characteristics. However, despite the progress made in modifying hydrogels for enhanced bioactivation, cell survival and tissue formation, little attention has so far been paid to optimize hydrogels for the physico-chemical demands of the biofabrication process. The resulting lack of hydrogel bioinks have been identified as one major hurdle for a more rapid progress of the field. In this review we summarize and focus on the deposition process, the parameters and demands of hydrogels in biofabrication, with special attention to robotic dispensing as an approach that generates constructs of clinically relevant dimensions. We aim to highlight this current lack of effectual hydrogels within biofabrication and initiate new ideas and developments in the design and tailoring of hydrogels. The successful development of a "printable" hydrogel that supports cell adhesion, migration, and differentiation will significantly advance this exciting and promising approach for tissue engineering.

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Topics: Biofabrication (70%), Self-healing hydrogels (56%), 3D bioprinting (52%)

1,180 Citations


Open accessJournal ArticleDOI: 10.1038/SREP00001
Upasana Ray1, Saumitra Das1Institutions (1)
14 Jun 2011-Scientific Reports
Abstract: HCV NS3 protein plays a central role in viral polyprotein processing and RNA replication. We demonstrate that the NS3 protease (NS3(pro)) domain alone can specifically bind to HCV-IRES RNA, predominantly in the SLIV region. The cleavage activity of the NS3 protease domain is reduced upon HCV-RNA binding. More importantly, NS3(pro) binding to the SLIV hinders the interaction of La protein, a cellular IRES-trans acting factor required for HCV IRES-mediated translation, resulting in inhibition of HCV-IRES activity. Although overexpression of both NS3(pro) as well as the full length NS3 protein decreased the level of HCV IRES mediated translation, replication of HCV replicon RNA was enhanced significantly. These observations suggest that the NS3(pro) binding to HCV IRES reduces translation in favor of RNA replication. The competition between the host factor (La) and the viral protein (NS3) for binding to HCV IRES might regulate the molecular switch from translation to replication of HCV.

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Topics: NS2-3 protease (64%), Internal ribosome entry site (60%), SeqA protein domain (57%) ... read more

859 Citations


Open accessJournal ArticleDOI: 10.3390/MA6041285
Jinchen Sun1, Huaping Tan1Institutions (1)
26 Mar 2013-Materials
Abstract: Alginate is a natural polysaccharide exhibiting excellent biocompatibility and biodegradability, having many different applications in the field of biomedicine Alginate is readily processable for applicable three-dimensional scaffolding materials such as hydrogels, microspheres, microcapsules, sponges, foams and fibers Alginate-based biomaterials can be utilized as drug delivery systems and cell carriers for tissue engineering Alginate can be easily modified via chemical and physical reactions to obtain derivatives having various structures, properties, functions and applications Tuning the structure and properties such as biodegradability, mechanical strength, gelation property and cell affinity can be achieved through combination with other biomaterials, immobilization of specific ligands such as peptide and sugar molecules, and physical or chemical crosslinking This review focuses on recent advances in the use of alginate and its derivatives in the field of biomedical applications, including wound healing, cartilage repair, bone regeneration and drug delivery, which have potential in tissue regeneration applications

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Topics: Bone regeneration (55%), Tissue engineering (54%), Biocompatibility (53%) ... read more

766 Citations


Journal ArticleDOI: 10.1038/NMAT3606
Hiroaki Onoe1, Teru Okitsu1, Akane Itou1, Midori Kato-Negishi1  +9 moreInstitutions (1)
01 Jun 2013-Nature Materials
Abstract: Artificial reconstruction of fibre-shaped cellular constructs could greatly contribute to tissue assembly in vitro. Here we show that, by using a microfluidic device with double-coaxial laminar flow, metre-long core-shell hydrogel microfibres encapsulating ECM proteins and differentiated cells or somatic stem cells can be fabricated, and that the microfibres reconstitute intrinsic morphologies and functions of living tissues. We also show that these functional fibres can be assembled, by weaving and reeling, into macroscopic cellular structures with various spatial patterns. Moreover, fibres encapsulating primary pancreatic islet cells and transplanted through a microcatheter into the subrenal capsular space of diabetic mice normalized blood glucose concentrations for about two weeks. These microfibres may find use as templates for the reconstruction of fibre-shaped functional tissues that mimic muscle fibres, blood vessels or nerve networks in vivo.

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608 Citations