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. 2019 Feb;71(1):261-266.
doi: 10.1007/s10616-018-0281-x. Epub 2019 Jan 1.

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

Sara Rastgar  1 , Rashid Alijani Ardeshir  1 , Abdolali Movahedinia  1   2 , Ebrahim Zabihi  3 , Amir Parviz Salati  4 , Negin Salamat  1
Affiliations

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

Sara Rastgar et al. Cytotechnology. 2019 Feb.

Abstract

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 aggregates, respectively. Also, grass carp heart explant had high potential in regeneration (especially in young fish) and thus high feasibility to generate stable long-term cultures. In general, it seems that explant culture of heart from grass carp can be considered as a promising tool in heart research area.

Keywords: Cardiac primary cells; Explant culture; Grass carp; Heart; Temperature.

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Conflict of interest statement

The authors declare no competing or financial interests.

Figures

Fig. 1
Fig. 1
Explant culture of heart from grass carp after 3–7 days. a Heart explant culture from group 1 after 3 days. Epithelium and fibroblast like cells are observed. b Heart explant culture from group 2 after 3 days. Most fibroblast like cells regularly and densely grew. c Heart explant culture from group 3 after 5 days. Fibroblast like cells are observed around the heart tissue. The rounded and detached cells (arrows) are dead. d Heart explant culture from group 4 after 7 days. Fibroblast cells grew in low density with increased number of dead cells (arrows)
Fig. 2
Fig. 2
Effect of temperature on beat frequency of spontaneously contracting heart cell aggregates from explant culture of heart from grass carp in different age groups. Different letters show significant difference (P < 0.05) in each group
Fig. 3
Fig. 3
Effect of FBS concentrations on beat frequency of spontaneously contracting cell aggregates derived from grass carp heart in different age groups. Different letters show significant difference (P < 0.05) in each group
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