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Establishing Sustainable Cell Lines of a Coral, Acropora tenuis
Marine Biotechnology ( IF 3 ) Pub Date : 2021-04-26 , DOI: 10.1007/s10126-021-10031-w
Kaz Kawamura 1 , Koki Nishitsuji 2 , Eiichi Shoguchi 2 , Shigeki Fujiwara 1 , Noriyuki Satoh 2
Affiliation  

Planula larvae of the scleractinian coral, Acropora tenuis, consist of elongated ectodermal cells and developing inner endodermal cells. To establish in vitro cell lines for future studies of cellular and developmental potential of coral cells, larvae were successfully dissociated into single cells by treating them with a tissue dissociation solution consisting of trypsin, EDTA, and collagenase. Brown-colored cells, translucent cells, and pale blue cells were the major components of dissociated larvae. Brown-colored cells began to proliferate transiently in the culture medium that was devised for the coral, while translucent cells and pale blue cells decreased in number about 1 week after cell dissociation. In addition, when a modular protease, plasmin, was added to the cell culture medium, brown-colored cells extended pseudopodia and assumed amorphous shapes. They then continued to proliferate in clumps for more than 6 months with a doubling time of approximately 4–5 days. From 3 weeks of cell culture onward, brown-colored cells often aggregated and exhibited morphogenesis-like behavior to form flat sheets, and blastula-like clusters or gastrula-like spheres. Single cells or cell-clusters of the cell lines were analyzed by RNA-seq. This analysis showed that genes expressed in these cells in vitro were A. tenuis genes. Furthermore, each cell line expressed a specific set of genes, suggesting that their properties include gastroderm, secretory cells, undifferentiated cells, neuronal cells, and epidermis. All cell properties were maintained stably throughout successive cell cultures. These results confirm the successful establishment of a coral in vitro cell line.



中文翻译:

建立珊瑚、Acropora tenuis 的可持续细胞系

scleractinian 珊瑚的 Planula 幼虫,Acropora tenuis,由拉长的外胚层细胞和发育中的内胚层细胞组成。为了建立体外细胞系,用于未来研究珊瑚细胞的细胞和发育潜力,通过用由胰蛋白酶、EDTA 和胶原酶组成的组织解离溶液处理幼虫,成功地将幼虫解离成单个细胞。棕色细胞、半透明细胞和淡蓝色细胞是离解幼虫的主要成分。棕色细胞在为珊瑚设计的培养基中开始短暂增殖,而半透明细胞和淡蓝色细胞在细胞分离后约 1 周数量减少。此外,当向细胞培养基中加入模块化蛋白酶纤溶酶时,棕色细胞会扩展伪足并呈现无定形形状。然后它们继续成团增殖超过 6 个月,倍增时间约为 4-5 天。从细胞培养 3 周起,棕色细胞经常聚集并表现出类似形态发生的行为,形成平板、囊胚样簇或原肠胚样球体。通过 RNA-seq 分析细胞系的单个细胞或细胞簇。该分析表明,这些细胞在体外表达的基因是A.弱基因。此外,每个细胞系都表达一组特定的基因,表明它们的特性包括胃胚、分泌细胞、未分化细胞、神经元细胞和表皮。在连续的细胞培养过程中,所有细胞特性都保持稳定。这些结果证实了珊瑚体外细胞系的成功建立。

更新日期:2021-04-26
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