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In Vitro Culture and Characterization of Testis-Derived Cells from Clarias magur (Hamilton, 1822)

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

A cell-based model offers many potential applications in biology. In vitro culture of testis-originated cells of Clarias magur was carried out using explant method. The cells were grown in different media compositions (including fetal bovine serum), concentrations, and temperatures. The developed culture was grown up to five passages. Leibovitz-15 (L-15) media with 20% FBS was found suitable, and culture conditions between 25 and 28 °C temperatures supported cell growth. The study is further focused on evaluation of the requirement of the other supplements along with the favorable basal media for growth and maintenance of testis-derived cell culture of C. magur. Epithelial-like cells in morphology were observed which were further confirmed with cell-specific markers. Chromosome observation revealed diploid number of 50 chromosomes as reported for the species. The species specificity of cultured cells was confirmed by amplification of two universal markers COI and 16S rRNA. The present study can be useful in the development of testicular cell line of C. magur.

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Acknowledgements

The authors are grateful to the Director, ICAR-NBFGR, Lucknow, for providing necessary facilities to carry out the present work. The financial support provided by Department of Biotechnology (DBT), New Delhi, vide Sanction Order No. BT/PR13908/AAQ/3/733/2015 dated 10.03.2017 is duly acknowledged.

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Authors and Affiliations

  1. Molecular Biology and Biotechnology Division, ICAR- National Bureau of Fish Genetic Resources, Lucknow, Uttar Pradesh, 226 002, India

    Neha Singh, Murali S, Basdeo Kushwaha, Shreya Srivastava, Akhilesh Kumar Mishra & Ravindra Kumar

  2. AMITY Institute of Biotechnology, AMITY University, Lucknow Campus, Lucknow, Uttar Pradesh, 226 028, India

    Neha Singh & J. K. Srivastava

Authors
  1. Neha Singh
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  2. Murali S
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  3. Basdeo Kushwaha
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  4. J. K. Srivastava
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  5. Shreya Srivastava
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  6. Akhilesh Kumar Mishra
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  7. Ravindra Kumar
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Correspondence to Basdeo Kushwaha.

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The authors declare that they have no conflict of interest to publish this manuscript.

Ethical Standards

Experiments under this study were conducted in accordance with the guidelines and regulations of Animal Ethics Committee which was constituted under guidance of Committee for the purpose of Control and Supervision of the Experiments on Animals (CPCSEA) under the provisions of the Prevention of Cruelty to Animals Act 1960, a Legislation of Ministry of Environment & Forests and Climate Change, Government of India. It is further mentioned that the experimental protocols were approved by the Institute Animal Ethics Committee of ICAR-National Bureau of Fish Genetic Resources, Lucknow, India.

Additional information

Significance statement In vitro culture of testis-originated cells of Clarias magur was carried out using explant method. The present study can be useful in the development of testicular cell line of C. magur.

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Singh, N., S, M., Kushwaha, B. et al. In Vitro Culture and Characterization of Testis-Derived Cells from Clarias magur (Hamilton, 1822). Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 89, 1017–1025 (2019). https://doi.org/10.1007/s40011-018-1020-y

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  • DOI: https://doi.org/10.1007/s40011-018-1020-y

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