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Establishment condition and characterization of heart-derived cell culture in Siberian sturgeon (Acipenser baerii)

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Abstract

This study was conducted to establish the efficient condition for stable derivation of heart-derived cell culture in Siberian sturgeon (Acipenser baerii). Three factors including isolation methods, cell densities in initial seeding, and basal media were evaluated for the derivation of heart-derived cell culture. As the results, enzymatic isolation was more efficient than mechanical isolation in both cell retrieval and further culture. Total 48 trials of culture employing low and middle cell densities of less than 5.5 × 104 cells/cm2 in initial seeding did not induce cell survivals (0%, 0/48), but the trials in high cell density of more than 5.5 × 105 cells/cm2 could induce cell survival and primary cell attachment on the plate (88.9%, 24 in 27 trials). When all initially attached cell populations were continuously cultured in two different media, only five cell populations that were enzymatically isolated and cultured under Leibovitz’s L-15 medium could grow up to more than 40th subculture. Each cell population was stably cultured according to its own growth rate and all showed normal diploid DNA contents. Two morphologically different cell types that has an elongated shape or a round shape were identified in culture, which was subsequently identified that two cell types are considered as a fibroblast (an elongated shape) and a vascular endothelial cell (a round shape) on the basis of the results of gene and protein expression analysis. Additionally, the sufficient number of viable cells could be successfully retrieved after freezing and thawing from all five cell populations suggesting the feasibility of long-term cryopreservation of the cells. The data and cells obtained from this study will contribute to development of in vitro model for basic biological studies using sturgeon species.

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Acknowledgment

This work was supported by a Research Grant of Pukyong National University (C-D-2013-0578).

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Authors and Affiliations

  1. Department of Fisheries Biology, Pukyong National University, Busan, Korea

    Min Sung Kim, Yoon Kwon Nam, Chulhong Park & Seung Pyo Gong

  2. Department of Marine Biomaterials and Aquaculture, Pukyong National University, Busan, Korea

    Yoon Kwon Nam & Seung Pyo Gong

  3. Department of Marine Biology, Pukyong National University, Busan, Korea

    Hyun-Woo Kim

  4. Stem Cell and Bioevaluation, WCU Biomodulation Program, Seoul National University, Seoul, Korea

    Jiyeon Ahn & Jeong Mook Lim

  5. Laboratory of Cell Biotechnology, Department of Marine Biomaterials and Aquaculture, College of Fisheries Science, Pukyong National University, Busan, Korea

    Seung Pyo Gong

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  1. Min Sung Kim
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  4. Hyun-Woo Kim
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Correspondence to Seung Pyo Gong.

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Editor: T. Okamoto

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Kim, M.S., Nam, Y.K., Park, C. et al. Establishment condition and characterization of heart-derived cell culture in Siberian sturgeon (Acipenser baerii). In Vitro Cell.Dev.Biol.-Animal 50, 909–917 (2014). https://doi.org/10.1007/s11626-014-9793-7

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  • DOI: https://doi.org/10.1007/s11626-014-9793-7

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