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Establishment of a testis cell line from Clarias magur: a potential resource for in-vitro applications

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Abstract

We developed a gonad cell line, CMgT-1, from the testes of Clarias magur in L-15 media with 20% FBS at 28 °C temperature and sub-cultured it more than 50 times. The origin of the cell line was validated using two universal markers, viz. cytochrome oxidase c subunit I and 16S rRNA genes. Cytogenetic analysis of the cells at 32nd passage revealed diploid chromosome number of 50. Immuno-phenotyping assay confirmed an epithelial-like cell type. The cell line was tested for mycoplasma contamination and found to be negative. The cells were successfully transfected with a GFP reporter gene that indicated that the cell line could be utilized in gene expression studies. The cell line was successfully utilized for evaluating cytotoxic effects of heavy metals, viz. potassium dichromate, mercury chloride and sodium (meta) arsenite, which reflects its potential applications in toxicity studies. CMgT-1 cell line will serve as a valuable resource for cytotoxicity and other in vitro studies.

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Availability of data and material (data transparency)

The deposited testis cell line CMgT-1 (Accession code: NRFC062) is presently available at National Repository of Fish Cell Lines (NRFC) at ICAR-National Bureau of Fish Genetic Resources (NBFGR), Lucknow, India for distribution to researchers for R&D works.

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Acknowledgements

The authors are thankful to the Director, ICAR-NBFGR, Lucknow, for support and providing the necessary facilities to carry out this work. The financial assistance granted by the Department of Biotechnology, Government of India, New Delhi, India, vide Sanction Order No. BT/PR13908/AAQ/3/733/2015 is duly acknowledged. The first author is also thankful to Dr. M. Goswami, Principal Scientist, ICAR-CIFE, Mumbai and former faculty of ICAR-NBFGR, Lucknow for his support.

Funding

The present work was carried out under a research project funded by the Department of Biotechnology, Government of India, New Delhi, India, vide Sanction Order No. BT/PR13908/AAQ/3/733/2015 dated May 10, 2017.

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Authors and Affiliations

  1. ICAR-National Bureau of Fish Genetic Resources, Canal Ring Road, P.O.-Dilkusha, Lucknow, Uttar Pradesh, 226002, India

    Neha Singh, Pankaj Soni, Basdeo Kushwaha, Murali Sanjeev Kumar, Shreya Srivastava, Akhilesh Kumar Mishra & Ravindra Kumar

  2. Amity Institute of Biotechnology, Amity University, Lucknow, Uttar Pradesh, 226010, India

    Neha Singh & J. K. Srivastava

Authors
  1. Neha Singh
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  2. Pankaj Soni
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  3. Basdeo Kushwaha
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  4. Murali Sanjeev Kumar
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  5. J. K. Srivastava
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  6. Shreya Srivastava
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  7. Akhilesh Kumar Mishra
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  8. Ravindra Kumar
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Contributions

NS: Conducted main experimentation, culture setup and manuscript draft preparation, PS: Assisted in cell culture and immuno-phenotyping, BK: Conceptualization, planning, supervision and manuscript preparation, MSK: Experiment designing and cell culture characterization, JKS: Supervision and manuscript editing, SS: Explant preparation and karyotyping, AKM: Sequencing and characterization, RK: Manuscript editing and proof reading.

Corresponding author

Correspondence to Basdeo Kushwaha.

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Conflict of interest

The authors declare that there is no conflict of interest in the present study.

Ethics approval

The guidelines issued by the Government of India for control and supervision of experiments on animals was followed strictly while handling the fish specimen. The present study was also approved by the Institute Animal Ethics Committee (IAEC) constituted by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), New Delhi, India.

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Corresponding Editor : Rabindra Nath Chatterjee

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Singh, N., Soni, P., Kushwaha, B. et al. Establishment of a testis cell line from Clarias magur: a potential resource for in-vitro applications. Nucleus 64, 211–217 (2021). https://doi.org/10.1007/s13237-020-00345-w

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  • DOI: https://doi.org/10.1007/s13237-020-00345-w

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