Production of zebrafish cardiospheres and cardiac progenitor cells in vitro and three‐dimensional culture of adult zebrafish cardiac tissue in scaffolds

Zeng, Wendy Rong; Beh, Siew‐Joo; Bryson-Richardson, Robert; Doran, Pauline M.

doi:10.1002/bit.26331

Cited by 7 publications

(5 citation statements)

References 18 publications

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“…In order to resolve this problem, primary adult cardiomyocytes ( Sander et al, 2013 ) and cardiospheres ( Zeng et al, 2017 ) from the zebrafish heart have been cultured successfully. Yet these systems lack the context of an injury, maturity of cardiomyocytes and the complexity of other cell types present in the heart.…”

Section: Introductionmentioning

confidence: 99%

Live imaging of adult zebrafish cardiomyocyte proliferation ex vivo

et al. 2021

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Zebrafish are excellent at regenerating their heart by reinitiating proliferation in pre-existing cardiomyocytes. Studying how zebrafish achieve this holds great potential in developing new strategies to boost mammalian heart regeneration. Nevertheless, the lack of appropriate live imaging tools for the adult zebrafish heart has limited detailed studies into the dynamics underlying cardiomyocyte proliferation. Here, we address this by developing a system in which cardiac slices of the injured zebrafish heart are cultured ex vivo for several days while retaining key regenerative characteristics including cardiomyocyte proliferation. In addition, we show that the cardiac slice culture system is compatible with live timelapse imaging and allows manipulation of regenerating cardiomyocytes with drugs that normally would have toxic effects that prevent its use. Finally, we use the cardiac slices to demonstrate that adult cardiomyocytes with fully assembled sarcomeres can partially disassemble their sarcomeres in a calpain and proteasome dependent manner to progress through nuclear division and cytokinesis. In conclusion, we have developed a cardiac slice culture system, which allows imaging of native cardiomyocyte dynamics in real time to discover cellular mechanisms during heart regeneration.

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Section: Introductionmentioning

confidence: 99%

Live imaging of adult zebrafish cardiomyocyte proliferation ex vivo

et al. 2021

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“…This alteration was more constant in the fish with higher age (2 and 3 years old). An increase in FBS concentration may introduce trophic factors that are essential for fish cardiac cell proliferation (Zeng et al 2017) and the fish with high age are more sensitive to this alteration compared to low age fish. Previous study on zebra fish showed that the increase in FBS concentrations led to cardiac cell proliferation and development (Zeng et al 2017).…”

Section: Discussionmentioning

confidence: 99%

“…An increase in FBS concentration may introduce trophic factors that are essential for fish cardiac cell proliferation (Zeng et al 2017) and the fish with high age are more sensitive to this alteration compared to low age fish. Previous study on zebra fish showed that the increase in FBS concentrations led to cardiac cell proliferation and development (Zeng et al 2017). Also, the present study showed that contraction frequencies over time were different (data not showed), that may be due to the change in the supply of growth factors as it occurs when the medium is changed (Grunow et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Spontaneously contracting cell aggregates derived from grass carp heart as a promising tool in in vitro heart research

et al. 2019

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Regarding challenges in isolation and maintenance of cultured heart cells, introduction of new in vitro heart model that is stable and exhibits long-term spontaneously contracting cell aggregates (SCCs), whose electrophysiological properties are comparable to the human heart, is required. This research aimed to establish cardiac primary cells and to evaluate the effects of culture conditions. Also the effect of fish age on beating SCC and cardiac cell morphology were studied. Twelve healthy grass carps (Ctenopharyngodon idella) were divided into four groups based on their age. Non-enzymatic explant culture was used and heart explants were incubated at 21-31°C for 60 days. After proliferation of the cardiac primary cells, changes in their morphology and their beatings were recorded. The results showed that the explants derived from different age of grass carp fish are fully viable and proliferative with formation of SCC for a long period of time. By increasing the number of adhered cells, the cardiac primary cells became more flat and elongated. Increasing the medium temperature and fetal bovine serum concentration in culture medium led to decline and enhancement in beat frequencies of heart cell Electronic supplementary material The online version of this article (

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“…Zebrafish are tropical fish that have recently emerged as a new and attractive model for studying physiological and pathological processes, and this is because of its high similarity to the human genome, transparent embryos and suitable for genetic manipulation 6,7 . The zebrafish animal model has been established as a disease model for cardiovascular diseases, and this is because of the fact that adult humans suffer from a high rate of cardiovascular diseases 8 . To date, there are several challenges posed in establishing the heart disease models in zebrafish, limiting their use in this field 9 .…”

Section: Introductionmentioning

confidence: 99%

Effect of adenosine monophosphate‐activated protein kinase–p53–Krüppel‐like factor 2a pathway in hyperglycemia‐induced cardiac remodeling in adult zebrafish

Wang

Luo

et al. 2020

J of Diabetes Invest

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Aims/Introduction Diabetic cardiomyopathy is a type of myocardial disease. It causes left ventricular hypertrophy, followed by diastolic and systolic dysfunction, eventually leading to congestive heart failure. However, the underlying mechanism still requires further elucidation. Materials and Methods A high‐glucose zebrafish model was constructed by administering streptozocin intraperitoneally to enhance the development of cardiomyopathy and then treated with adenosine monophosphate‐activated protein kinase (AMPK) activator. Cardiac structure and function, and protein and gene expression were then analyzed. Cardiomyocytes (CMs) culture in vitro using lentivirus were used for detection of AMPK, p53 and Krüppel‐like factor 2a (klf2a) gene expression. Results In the hyperglycemia group, electrocardiogram findings showed arrhythmia, echocardiography results showed heart enlargement and dysfunction, and many differences, such as increased apoptosis and myocardial fiber loss, were observed. The phospho‐AMPK and klf2a expression were downregulated, and p53 expression was upregulated. Activation of phospho‐AMPK reduced p53 and increased klf2a expression, alleviated apoptosis in CMs and improved cardiac function in the hyperglycemic zebrafish. In vitro knockdown system of AMPK, p53 and klf2a using lentivirus illustrated an increased p53 expression and decreased klf2a expression in CMs by inhibiting AMPK. Repression of p53 and upregulation of klf2a expression were observed, but no changes in the expression of AMPK and its phosphorylated type. Conclusions In the model of streptozocin‐induced hyperglycemia zebrafish, the reduction of phosphorylated AMPK increased p53, which led to KLF2a decrease to facilitate apoptosis of CMs, inducing the cardiac remodeling and cardiac dysfunction. These results can be reversed by AMPK activator, which means the AMPK–p53–klf2a pathway might be a potential target for diabetic cardiomyopathy intervention.

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Production of zebrafish cardiospheres and cardiac progenitor cells in vitro and three‐dimensional culture of adult zebrafish cardiac tissue in scaffolds

Cited by 7 publications

References 18 publications

Live imaging of adult zebrafish cardiomyocyte proliferation ex vivo

Live imaging of adult zebrafish cardiomyocyte proliferation ex vivo

Spontaneously contracting cell aggregates derived from grass carp heart as a promising tool in in vitro heart research

Effect of adenosine monophosphate‐activated protein kinase–p53–Krüppel‐like factor 2a pathway in hyperglycemia‐induced cardiac remodeling in adult zebrafish

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