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

  • Min Sung Kim
  • Yoon Kwon Nam
  • Chulhong Park
  • Hyun-Woo Kim
  • Jiyeon Ahn
  • Jeong Mook Lim
  • Seung Pyo GongEmail author


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.


Siberian sturgeon Acipenser baerii Cell culture Heart tissue 



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


  1. Beauchamp NJ, van Achterberg TA, Engelse MA, Pannekoek H, de Vries CJ (2003) Gene expression profiling of resting and activated vascular smooth muscle cells by serial analysis of gene expression and clustering analysis. Genomics 82:288–299PubMedCrossRefGoogle Scholar
  2. Bemis WE, Findeis EK, Grande L (1997) An overview of Acipenseriformes. Environ Biol Fishes 48:25–71CrossRefGoogle Scholar
  3. Bradshaw AD, Francki A, Motamed K, Howe C, Sage EH (1999) Primary mesenchymal cells isolated from SPARC-null mice exhibit altered morphology and rates of proliferation. Mol Biol Cell 10:1569–1579PubMedCentralPubMedCrossRefGoogle Scholar
  4. Chen SL, Ren GC, Sha ZX, Shi CY (2004) Establishment of a continuous embryonic cell line from Japanese flounder Paralichthys olivaceus for virus isolation. Dis Aquat Organ 60:241–246PubMedCrossRefGoogle Scholar
  5. Ciba P, Schicktanz S, Anders E, Siegl E, Stielow A, Klink E, Kruse C (2008) Long-term culture of a cell population from Siberian sturgeon (Acipenser baerii) head kidney. Fish Physiol Biochem 34:367–372PubMedCrossRefGoogle Scholar
  6. Freshney RI (2010) Culture of animal cells: a manual of basic technique and specialized applications. Wiley-Liss, New YorkCrossRefGoogle Scholar
  7. Grunow B, Noglick S, Kruse C, Gebert M (2011) Isolation of cells from Atlantic sturgeon Acipenser oxyrinchus oxyrinchus and optimization of culture conditions. Aquat Biol 14:67–75CrossRefGoogle Scholar
  8. Grzelak A, Rychlik B, Bartosz G (2001) Light-dependent generation of reactive oxygen species in cell culture media. Free Radic Biol Med 30:1418–1425PubMedCrossRefGoogle Scholar
  9. Hedrick RP, McDowell T, RosemarkR AD, Lannan CN (1991) Two cell lines from white sturgeon. Trans Am Fish Soc 120:528–534CrossRefGoogle Scholar
  10. Hong N, Schartl M, Hong Y (2014) Derivation of stable zebrafish ES-like cells in feeder-free culture. Cell Tissue Res. doi: 10.1007/s00441-014-1882-0 Google Scholar
  11. Icardo JM, Colvee E, Cerra MC, Tota B (2002a) Structure of the conus arteriosus of the sturgeon (Acipenser naccarii) heart. I: the conus valves and the subendocardium. Anat Rec 267:17–27PubMedCrossRefGoogle Scholar
  12. Icardo JM, Colvee E, Cerra MC, Tota B (2002b) The structure of the conus arteriosus of the sturgeon (Acipenser naccarii) heart: II. The myocardium, the subepicardium, and the conus-aorta transition. Anat Rec 268:388–398PubMedCrossRefGoogle Scholar
  13. Krieger J, Fuerst PA (2002) Evidence of multiple alleles of the nuclear 18S ribosomal RNA gene in sturgeon (Family: Acipenseridae). J Appl Ichthyol 18:290–297CrossRefGoogle Scholar
  14. Lee D, Kim MS, Nam YK, Kim DS, Gong SP (2013) Establishment and characterization of permanent cell lines from Oryzias dancena embryos. Fish Aquat Sci 16:177–185Google Scholar
  15. Li MF, Marrayatt V, Annand C, Odense P (1985) Fish cell culture: two newly developed cell lines from Atlantic sturgeon (Acipenser oxyrhynchus) and guppy (Poecilia reticulata). Can J Zool 63:2867–2874CrossRefGoogle Scholar
  16. Li Z, Mericskay M, Agbulut O, Butler-Browne G, Carlsson L, Thornell LE, Babinet C, Paulin D (1997) Desmin is essential for the tensile strength and integrity of myofibrils but not for myogenic commitment, differentiation, and fusion of skeletal muscle. J Cell Biol 139:129–144PubMedCentralPubMedCrossRefGoogle Scholar
  17. Limoli CL, Rola R, Giedzinski E, Mantha S, Huang TT, Fike JR (2004) Cell-density-dependent regulation of neural precursor cell function. Proc Natl Acad Sci USA 101:16052–16057PubMedCentralPubMedCrossRefGoogle Scholar
  18. Liu T, Guevara OE, Warburton RR, Hill NS, Gaestel M, Kayyali US (2010) Regulation of vimentin intermediate filaments in endothelial cells by hypoxia. Am J Physiol Cell Physiol 299:C363–C373PubMedCentralPubMedCrossRefGoogle Scholar
  19. Ohgushi M, Matsumura M, Eiraku M, Murakami K, Aramaki T, Nishiyama A, Muguruma K, Nakano T, Suga H, Ueno M, Ishizaki T, Suemori H, Narumiya S, Niwa H, Sasai Y (2010) Molecular pathway and cell state responsible for dissociation-induced apoptosis in human pluripotent stem cells. Cell Stem Cell 7:225–239PubMedCrossRefGoogle Scholar
  20. Parkinson DB, Dong Z, Bunting H, Whitfield J, Meier C, Marie H, Mirsky R, Jessen KR (2001) Transforming growth factor beta (TGFbeta) mediates Schwann cell death in vitro and in vivo: examination of c-Jun activation, interactions with survival signals, and the relationship of TGFbeta-mediated death to Schwann cell differentiation. J Neurosci 21:8572–8585PubMedGoogle Scholar
  21. Pikitch EK, Doukakis P, Lauck L, Chakrabarty P, Erickson DL (2005) Status, trends and management of sturgeon and paddlefish fisheries. Fish Fish 6:233–265CrossRefGoogle Scholar
  22. Platet N, Liu SY, Atifi ME, Oliver L, Vallette FM, Berger F, Wion D (2007) Influence of oxygen tension on CD133 phenotype in human glioma cell cultures. Cancer Lett 258:286–290PubMedCrossRefGoogle Scholar
  23. Portela VM, Zamberlam G, Price CA (2010) Cell plating density alters the ratio of estrogenic to progestagenic enzyme gene expression in cultured granulosa cells. Fertil Steril 93:2050–2055PubMedCrossRefGoogle Scholar
  24. Robles F, de la Herrán R, Ludwig A, Ruiz Rejón C, Ruiz Rejón M, Garrido-Ramos MA (2004) Evolution of ancient satellite DNAs in sturgeon genomes. Gene 338:133–142PubMedCrossRefGoogle Scholar
  25. Souders CA, Bowers SL, Baudino TA (2009) Cardiac fibroblast: the renaissance cell. Circ Res 105:1164–1176PubMedCentralPubMedCrossRefGoogle Scholar
  26. Spörl F, Wunderskirchner M, Ullrich O, Bömke G, Breitenbach U, Blatt T, Wenck H, Wittern KP, Schrader A (2010) Real-time monitoring of membrane cholesterol reveals new insights into epidermal differentiation. J Invest Dermatol 130:1268–1278PubMedCrossRefGoogle Scholar
  27. Vegusdal A, Ostbye TK, Tran TN, Gjøen T, Ruyter B (2004) Beta-oxidation, esterification, and secretion of radiolabeled fatty acids in cultivated Atlantic salmon skeletal muscle cells. Lipids 39:649–658PubMedCrossRefGoogle Scholar
  28. Wang D, Zhong L, Wei Q, Gan X, He S (2010) Evolution of MHC class I genes in two ancient fish, paddlefish (Polyodon spathula) and Chinese sturgeon (Acipenser sinensis). FEBS Lett 584:3331–3339Google Scholar
  29. Wang G, LaPatra S, Zeng L, Zhao Z, Lu Y (2003) Establishment, growth, cryopreservation and species of origin identification of three cell lines from white sturgeon, Acipenser transmontanus. Methods Cell Sci 25:211–220PubMedCrossRefGoogle Scholar
  30. Watson LR, Groff JM, Hedrick RP (1998) Replication and pathogenesis of white sturgeon iridovirus (WSIV) in experimentally infected white sturgeon Acipenser transmontanus juveniles and sturgeon cell lines. Dis Aquat Organ 32:173–184PubMedCrossRefGoogle Scholar
  31. Will MA, Clark NA, Swain JE (2011) Biological pH buffers in IVF: help or hindrance to success. J Assist Reprod Genet 28:711–724PubMedCentralPubMedCrossRefGoogle Scholar
  32. Wolters GH, Vos-Scheperkeuter GH, van Deijnen JH, van Schilfgaarde R (1992) An analysis of the role of collagenase and protease in the enzymatic dissociation of the rat pancreas for islet isolation. Diabetologia 35:735–742PubMedCrossRefGoogle Scholar
  33. Yi M, Hong N, Hong Y (2009) Generation of medaka fish haploid embryonic stem cells. Science 326:430–433PubMedCrossRefGoogle Scholar
  34. Zhou GZ, Gui L, Li ZQ, Yuan XP, Zhang QY (2008) Establishment of a Chinese sturgeon Acipenser sinensis tail-fin cell line and its susceptibility to frog iridovirus. J Fish Biol 73:2058–2067CrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2014

Authors and Affiliations

  • Min Sung Kim
    • 1
  • Yoon Kwon Nam
    • 1
    • 2
  • Chulhong Park
    • 1
  • Hyun-Woo Kim
    • 3
  • Jiyeon Ahn
    • 4
  • Jeong Mook Lim
    • 4
  • Seung Pyo Gong
    • 1
    • 2
    • 5
    Email author
  1. 1.Department of Fisheries BiologyPukyong National UniversityBusanKorea
  2. 2.Department of Marine Biomaterials and AquaculturePukyong National UniversityBusanKorea
  3. 3.Department of Marine BiologyPukyong National UniversityBusanKorea
  4. 4.Stem Cell and Bioevaluation, WCU Biomodulation ProgramSeoul National UniversitySeoulKorea
  5. 5.Laboratory of Cell Biotechnology, Department of Marine Biomaterials and Aquaculture, College of Fisheries SciencePukyong National UniversityBusanKorea

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