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Development of a zebrafish spleen cell line, ZSSJ, and its growth arrest by gamma irradiation and capacity to act as feeder cells

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Abstract

A zebrafish spleen cell line, ZSSJ, was developed and its growth arrest by gamma radiation determined and its capacity to stimulate the proliferation of the zebrafish blastula cell line, ZEB2J, measured. ZSSJ was initiated by explant outgrowth, grew adherent with mainly an epithelial-like morphology, and stained strongly for alkaline phosphatase. ZSSJ was not only grown in L-15 with 15% fetal bovine serum at 26°C to 28°°C but also grew at room temperature. Cultures of ZSSJ have undergone approximately 40 population doublings, had few cells staining for b-galactosidase activity, which is commonly present in senescent cultures, and many cells with an aneuploid karyotype, which is frequently associated with immortalization. ZSSJ growth was arrested by 30 to 50 Gy of g-irradiation, whereas after 20 Gy, some slight growth was observed. By contrast, growth of the rainbow trout spleen stromal cell line, RTS34st, which has been used as a feeder for zebrafish ES cell cultures, was arrested completely by 20 Gy. In cocultures, nongrowth-arrested ZSSJ stimulated ZEB2J proliferation better than growth-arrested ZSSJ and better than RTS34st. ZSSJ should be useful as a feeder cell line for zebrafish ES cell cultures.

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Acknowledgments

This research was supported by grants from the U. S. National Institute of Health (NIH GM069383) and from the Natural Sciences and Engineering Research Council of Canada (NSERC). The authors thank Rob Barnett of the Grand River Hospital for help with the gamma irradiation.

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Authors and Affiliations

  1. Department of Biology, University of Waterloo, 200 University Ave., Waterloo, ON, Canada, N2L 3G1

    J. G. Xing, W. El-Sweisi & N. C. Bols

  2. Department of Biology, Wilfrid Laurier University, Waterloo, ON, Canada

    L. E. J. Lee

  3. Department of Animal Sciences, Purdue University, West Lafayette, IN, USA

    P. Collodi

  4. Department of Medical Physics & Applied Radiation Science, McMaster University, Hamilton, ON, Canada

    C. Seymour & C. Mothersill

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  1. J. G. Xing
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  2. W. El-Sweisi
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Correspondence to N. C. Bols.

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Editor: J. Denry Sato

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Xing, J.G., El-Sweisi, W., Lee, L.E.J. et al. Development of a zebrafish spleen cell line, ZSSJ, and its growth arrest by gamma irradiation and capacity to act as feeder cells. In Vitro Cell.Dev.Biol.-Animal 45, 163–174 (2009). https://doi.org/10.1007/s11626-008-9159-0

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  • DOI: https://doi.org/10.1007/s11626-008-9159-0

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