Generating an <i>In Vitro</i> 3D Cell Culture Model from Zebrafish Larvae for Heart Research

Grunow, Bianka; Mohamet, Lisa; Shiels, Holly A.

doi:10.1242/jeb.118224

Cited by 14 publications

(23 citation statements)

References 34 publications

(48 reference statements)

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“…In particular, fish heart models have been found to have a higher similarity to the characteristics of the human heart than the traditional murine models and have been therefore suggested as a functional substitute for in vitro toxicological testing [15,19–21,32]. …”

Section: Discussionmentioning

confidence: 99%

Atlantic salmon cardiac primary cultures: An in vitro model to study viral host pathogen interactions and pathogenesis

et al. 2017

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Development of Salmon Cardiac Primary Cultures (SCPCs) from Atlantic salmon pre-hatch embryos and their application as in vitro model for cardiotropic viral infection research are described. Producing SCPCs requires plating of trypsin dissociated embryos with subsequent targeted harvest from 24h up to 3 weeks, of relevant tissues after visual identification. SCPCs are then transferred individually to chambered wells for culture in isolation, with incubation at 15–22°. SCPCs production efficiency was not influenced by embryo’s origin (0.75/ farmed or wild embryo), but mildly influenced by embryonic developmental stage (0.3 decline between 380 and 445 accumulated thermal units), and strongly influenced by time of harvest post-plating (0.6 decline if harvested after 72 hours). Beating rate was not significantly influenced by temperature (15–22°) or age (2–4 weeks), but was significantly lower on SCPCs originated from farmed embryos with a disease resistant genotype (F = 5.3, p<0.05). Two distinct morphologies suggestive of an ex vivo embryonic heart and a de novo formation were observed sub-grossly, histologically, ultra-structurally and with confocal microscopy. Both types contained cells consistent with cardiomyocytes, endothelium, and fibroblasts. Ageing of SCPCs in culture was observed with increased auto fluorescence in live imaging, and as myelin figures and cellular degeneration ultra-structurally. The SCPCs model was challenged with cardiotropic viruses and both the viral load and the mx gene expression were measurable along time by qPCR. In summary, SCPCs represent a step forward in salmon cardiac disease research as an in vitro model that partially incorporates the functional complexity of the fish heart.

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Section: Discussionmentioning

confidence: 99%

Atlantic salmon cardiac primary cultures: An in vitro model to study viral host pathogen interactions and pathogenesis

et al. 2017

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“…While these genetic strategies are commonplace in mouse, similar tools are only just emerging in the zebrafish community (Ablain et al, 2015). Moreover, basic reagents such as antibodies and tools including cell culture techniques that are readily available to the research community for studying glial cell development in mouse are limited (though improving) for use in zebrafish (Staudt et al, 2015; Tapanes-Castillo et al, 2014; Hong et al, 2014; Grunow et al, 2015). …”

Section: Zebrafish and Mouse Advantages And Disadvantagesmentioning

confidence: 99%

The scales and tales of myelination: using zebrafish and mouse to study myelinating glia

Ackerman

Monk

2016

Brain Research

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Myelin, the lipid-rich sheath that insulates axons to facilitate rapid conduction of action potentials, is an evolutionary innovation of the jawed-vertebrate lineage. Research efforts aimed at understanding the molecular mechanisms governing myelination have primarily focused on rodent models; however, with the advent of the zebrafish model system in the late twentieth century, the use of this genetically tractable, yet simpler vertebrate for studying myelination has steadily increased. In this review, we compare myelinating glial cell biology during development and regeneration in zebrafish and mouse and enumerate the advantages and disadvantages of using each model to study myelination.

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“…To control the structure, morphology, and function of 3D cellular models, several strategies have been developed, including rotary cell cultures, microcarrier beads, gyratory shakers and roller tubes, spinner flask cultures, hanging drop method, liquid overlay cultures, and spontaneous cell aggregation methods (Figure d) . Another approach is to encapsulate or seed cells in/on biomaterial scaffolds, providing a controllable microenvironment for the cells.…”

Section: Introductionmentioning

confidence: 99%

Gelatin‐polysaccharide composite scaffolds for 3D cell culture and tissue engineering: Towards natural therapeutics

Afewerki

Sheikhi

Kannan

et al. 2018

Bioengineering & Transla Med

290

210

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Gelatin is a promising material as scaffold with therapeutic and regenerative characteristics due to its chemical similarities to the extracellular matrix (ECM) in the native tissues, biocompatibility, biodegradability, low antigenicity, cost‐effectiveness, abundance, and accessible functional groups that allow facile chemical modifications with other biomaterials or biomolecules. Despite the advantages of gelatin, poor mechanical properties, sensitivity to enzymatic degradation, high viscosity, and reduced solubility in concentrated aqueous media have limited its applications and encouraged the development of gelatin‐based composite hydrogels. The drawbacks of gelatin may be surmounted by synergistically combining it with a wide range of polysaccharides. The addition of polysaccharides to gelatin is advantageous in mimicking the ECM, which largely contains proteoglycans or glycoproteins. Moreover, gelatin–polysaccharide biomaterials benefit from mechanical resilience, high stability, low thermal expansion, improved hydrophilicity, biocompatibility, antimicrobial and anti‐inflammatory properties, and wound healing potential. Here, we discuss how combining gelatin and polysaccharides provides a promising approach for developing superior therapeutic biomaterials. We review gelatin–polysaccharides scaffolds and their applications in cell culture and tissue engineering, providing an outlook for the future of this family of biomaterials as advanced natural therapeutics.

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Generating an In Vitro 3D Cell Culture Model from Zebrafish Larvae for Heart Research

Cited by 14 publications

References 34 publications

Atlantic salmon cardiac primary cultures: An in vitro model to study viral host pathogen interactions and pathogenesis

Atlantic salmon cardiac primary cultures: An in vitro model to study viral host pathogen interactions and pathogenesis

The scales and tales of myelination: using zebrafish and mouse to study myelinating glia

Gelatin‐polysaccharide composite scaffolds for 3D cell culture and tissue engineering: Towards natural therapeutics

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