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Establishment and characterization of a cell line from the Chinese soft-shelled turtle Pelodiscus sinensis

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Abstract

The establishment and partial characterization of Pelodiscus sinensis continuous cell line is described here. A novel P. sinensis fibroblast cell line, designated PSF, was established from heart tissue by the semi-digestion explant culture technique. Since its initiation in July 2013, the cell line has been subcultured at 30°C in minimal essential medium (MEM) containing 15% (v/v) fetal bovine serum for more than 50 passages. The growth curve of the cell line revealed the population doubling time was 51.1 h. Karyotyping analysis indicated the modal chromosome number was 66, and no microbial contamination was detected. The PSF cell line produced significant fluorescent signals after transfection with plasmid pEGFP-C3. Analysis of mitochondrial cytochrome D-loop sequences revealed 96% identity among other Chinese turtle subspecies. Several cell line characterizations included morphological analysis and immunocytochemistry, which revealed the origin of the PSF cell line was fibroblast-like cells. Measurement of the isoenzymes lactic dehydrogenase and malic dehydrogenase showed no cross-contamination of this cell line with other species. This newly established cell line will be a valuable tool for transgenic and genetic manipulation studies and will act as an efficient instrument for studies of the viral diseases of the soft-shelled turtle.

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Acknowledgments

This study was funded by Zhejiang Provincial Project of Selective Breeding of Aquatic New Varieties (No. 2012C12907), Innovation Team Project of Ningbo Municipal Science and Technology Bureau (No. 2012B82016), and major agricultural projects of Ningbo Municipal Science and Technology Bureau (No. 2013C11011).

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Authors and Affiliations

  1. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, 201306, China

    Haijie Guo & Wei Tang

  2. Faculty of Biological and Environmental Sciences, Zhejiang Wanli University, No. 8, South Qian Hu Road, Ningbo, Zhejiang Province, 315100, China

    Haijie Guo, Zhaonan Xia, Wei Tang, Zhijuan Mao, Guoying Qian & Caisheng Wang

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  1. Haijie Guo
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  2. Zhaonan Xia
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  3. Wei Tang
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  4. Zhijuan Mao
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  5. Guoying Qian
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  6. Caisheng Wang
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Correspondence to Zhijuan Mao or Guoying Qian.

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Editor: Tetsuji Okamoto

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Guo, H., Xia, Z., Tang, W. et al. Establishment and characterization of a cell line from the Chinese soft-shelled turtle Pelodiscus sinensis . In Vitro Cell.Dev.Biol.-Animal 52, 673–682 (2016). https://doi.org/10.1007/s11626-016-0015-3

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  • DOI: https://doi.org/10.1007/s11626-016-0015-3

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