Rastgar Sara, Alijani Ardeshir Rashid, Movahedinia Abdolali, Zabihi Ebrahim, Salati Amir Parviz, Salamat Negin
Department of Marine Biology, Faculty of Marine Sciences, Khorramshahr University of Marine Science and Technology, P.O. Box 669, Khorramshahr, Iran.
Department of Marine Biology, Faculty of Marine Sciences, University of Mazandaran, Babolsar, Iran.
Cytotechnology. 2019 Feb;71(1):261-266. doi: 10.1007/s10616-018-0281-x. Epub 2019 Jan 1.
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.
关于培养心脏细胞的分离和维持方面的挑战,需要引入一种新的体外心脏模型,该模型稳定且能呈现长期自发收缩的细胞聚集体(SCCs),其电生理特性与人类心脏相当。本研究旨在建立心脏原代细胞并评估培养条件的影响。此外,还研究了鱼龄对跳动的SCC和心脏细胞形态的影响。将12条健康草鱼(Ctenopharyngodon idella)按年龄分为四组。采用非酶外植体培养法,将心脏外植体在21 - 31°C下孵育60天。在心脏原代细胞增殖后,记录其形态变化和跳动情况。结果表明,来自不同年龄草鱼的外植体完全有活力且能增殖,并长时间形成SCC。随着贴壁细胞数量增加,心脏原代细胞变得更加扁平且细长。提高培养基温度和培养基中胎牛血清浓度分别导致心脏细胞聚集体的跳动频率下降和增加。此外,草鱼心脏外植体具有很高的再生潜力(尤其是幼鱼),因此生成稳定长期培养物的可行性很高。总体而言,草鱼心脏的外植体培养似乎可被视为心脏研究领域一种有前景的工具。