Application of Novel Gill Cell Line from Lates calcarifer for Recognizing Metals Using Probes.

IF 3.4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biological Trace Element Research Pub Date : 2025-02-01 Epub Date: 2024-05-23 DOI:10.1007/s12011-024-04229-x

Sivaraj Mithra, Seepoo Abdul Majeed, Arni Mujthaba Aatif, Selvam Suryakodi, Abdul Nafeez Ahmed, Gani Taju, Mohamed Jaffer Abdul Wazith, Palsamy Ramesh Kumar, Azeez Sait Sahul Hameed

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Abstract

Lates calcarifer (Bloch) is a potential candidate fish species for culture in marine and brackishwater. A continuous gill cell line was derived from L. calcarifer by the explant culture method and has been passaged for 132 times, in Leibovitz's L-15 medium containing 10% fetal bovine serum (FBS) at 28 °C. The cells showed a rate of recovery between 90 and 95% after being successfully cryopreserved at various passage levels and formed monolayer in 2-3 days without any morphological changes. Immunophenotypic analysis of the SBG cell line revealed that they are of epithelial origin. Polymerase chain reaction assay using mitochondrial 12S rRNA primer specific to L. calcarifer was used to confirm the authenticity of the established gill cell line origin from seabass. The transfection efficiency was evaluated in Seabass Gill (SBG) cell line using pEGFP-N1 and Lipofectamine™ 3000. Transfection efficiency was found to be between 13 and 16%. The cytotoxicity of three different metal detecting probes was evaluated by MTT and Alamar blue assays to determine safe concentration. The result revealed that SBG cell line can be applied for recognition of metals using probes. The current study established, for the first time, a gill-derived cell line (SBG) from Lates calcarifer and its application for the detection of intracellular indium, mercury, and lutetium ions by specific fluorescent probes.

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利用探针识别金属的鳐鱼新型鳃细胞系的应用

Lates calcarifer (Bloch) 是一种可用于海水和咸水养殖的潜在候选鱼种。通过外植体培养法从鲈鱼中获得了连续鳃细胞系，并在含有 10%胎牛血清（FBS）的 Leibovitz's L-15 培养基中于 28 °C下进行了 132 次传代。在不同的传代水平上成功冷冻保存后，细胞的恢复率在 90% 到 95% 之间，并在 2-3 天内形成单层，没有发生任何形态变化。对 SBG 细胞系的免疫表型分析表明，它们属于上皮细胞。使用线粒体 12S rRNA 引物进行聚合酶链反应检测，以确认所建立的鳃细胞系来源于鲈鱼。使用 pEGFP-N1 和 Lipofectamine™ 3000 评估了海鲈鳃细胞系（SBG）的转染效率。结果发现转染效率在 13% 到 16% 之间。通过 MTT 和阿拉玛蓝检测法评估了三种不同金属检测探针的细胞毒性，以确定安全浓度。结果表明，SBG 细胞系可用于使用探针识别金属。本研究首次建立了钙鲈鳃源细胞系（SBG），并将其应用于利用特异性荧光探针检测细胞内铟、汞和镥离子。

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来源期刊

Biological Trace Element Research 生物-内分泌学与代谢

CiteScore

8.70

自引率

10.30%

发文量

459

审稿时长

2 months

期刊介绍： Biological Trace Element Research provides a much-needed central forum for the emergent, interdisciplinary field of research on the biological, environmental, and biomedical roles of trace elements. Rather than confine itself to biochemistry, the journal emphasizes the integrative aspects of trace metal research in all appropriate fields, publishing human and animal nutritional studies devoted to the fundamental chemistry and biochemistry at issue as well as to the elucidation of the relevant aspects of preventive medicine, epidemiology, clinical chemistry, agriculture, endocrinology, animal science, pharmacology, microbiology, toxicology, virology, marine biology, sensory physiology, developmental biology, and related fields.