Establishment of glass catfish (Kryptopterus bicirrhis) fin-derived cells

Jee Eun Han, Casiano H Choresca, Ok Jae Koo, Hyun Ju Oh, So Gun Hong, Ji Hyung Kim, Sang Phil Shin, Jin Woo Jun, Byeong Chun Lee, Se Chang Park
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引用次数: 12

Abstract

Genetically manipulated transparent animals were already explored in many species for in vivo study of gene expression, transplantation analysis and cancer biology. However, there are no reports about transparent animals as in vitro genetic resources. In the present study, fin-derived cells from glass catfish (Krytopterus bicirrhis), naturally transparent fish with a visible skeleton and internal organs, were isolated after culturing fin explants and characterized using cryopreservation and cell cycle analysis. The cells grew well in DMEM (Dulbecco's modified Eagle's medium) containing 1% (v/v) P/S (penicillin—streptomycin) and 10% (v/v) fetal bovine serum at 26°C and showed increased cryopreservation efficiency with the slow-freezing method in the presence of 15% dimethyl sulfoxide. In addition, cell cycle analysis was evaluated based on flow cytometric analysis, and culturing to confluence (>85%) was more effective for synchronizing cells at the G0/G1 stages than roscovitine treatment (<75%). This is the first report about cell isolation from transparent animals. The results from testing the cell's viability following cryopreservation and subjecting the cells to cycle analysis can be useful tools for genetic resource management.

Abstract Image

玻璃鲶鱼鱼鳍细胞的建立
基因操纵透明动物已经在许多物种中进行了基因表达、移植分析和癌症生物学的体内研究。然而,目前还没有透明动物作为体外遗传资源的报道。本研究从玻璃鲶鱼(Krytopterus bicirrhis)中分离出鳍源性细胞,并通过冷冻保存和细胞周期分析对其进行了鉴定。玻璃鲶鱼是一种天然透明鱼类,具有可见的骨骼和内部器官。细胞在含有1% (v/v) P/S(青霉素-链霉素)和10% (v/v)胎牛血清的DMEM (Dulbecco’S modified Eagle’S培养基)中生长良好,在含有15%二甲亚砜的慢速冷冻法下,细胞的低温保存效率提高。此外,基于流式细胞术分析对细胞周期分析进行了评估,在G0/G1期,融合培养(>85%)比罗斯科维汀处理(<75%)更有效地同步细胞。这是关于透明动物细胞分离的第一篇报道。低温保存后测试细胞活力并对细胞进行周期分析的结果可作为遗传资源管理的有用工具。
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