Abstract
Eleven cell lines were prepared from skin, snout, liver, kidney, lung, heart, brain, spleen, thyroid, urinary bladder, and periorbital soft tissue of a juvenile Hawaiian monk seal (Monachus schauinslandi). The cell grew at 37 °C in RPMI 1640 medium supplemented with 20% fetal bovine serum. These cell lines have been subcultured 11–27 times since their initiation in May 1997. Growth of the monk seal cells was serum-dependent and plating efficiencies ranged from 4–24%. These monk seal cells grew well in M199, L-15 and MEM commonly used for cultivation of animal and mammalian cells and retained 87% cell viability following storage for 2.5 years in liquid nitrogen. Karyotyping indicated that these monk seal-derived cell lines remained diploid with a chromosome count of 34 at their early passage (passage 9–13). These cell lines were tested for herpesvirus by polymerase chain reaction using degenerate oligonucleotide primers designed from the highly conserved region of herpesviral DNA polymerase gene and no specific detection occurred. These newly established cell lines are currently being used for the investigation of an eye disease occurring in captive monk seal pups in Oahu and will be available for future isolation and study of monk seal viruses.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alcorn DJ, Kam AKH (1986). Fatal shark attack on a Hawaiian monk seal (Monachus schauinslandi). Marine Mammal Science 2: 313-315.
Arnason U (1974). Comparative chromosome studies in pinnipedia. Hereditas 70: 113-118.
Arnason U (1977). The relationship between the four principal pinniped karyotypes. Hereditas 87: 227-242.
Arnason U, Bodin K, Gullberg A, Ledje, C, Mouchaty S (1995). A molecular view of pinniped relationships with particular emphasis on the true seals. J Mol Evol 40: 78-85.
Balazs GH, Whittow G (1979). First record of a tiger shark observed feeding on a Hawaiian monk seal. 'Elepaio 39: 107-109.
Banish LD, Gilmartin WG (1988). Hematology and serum chemistry of the young Hawaiian monk seal (Monachus schauinslandi). J Wild Dis 24: 225-230.
Banish LD, Gilmartin WG (1992). Pathological findings in the Hawaiian monk seal. J Wild Dis 28: 428-434.
Birkun A Jr (1996). Viruses of whales and dolphins. Microbiol Zool 58: 100-106.
Birkun A Jr, Kuiken T, Krivokhizhin S, Haines DM, Osterhaus AD, van de Bildt MW, Joiris CR, Siebert U (1999). Epixzootic of morbilliviral disease in common dolphins (Delphinus delphis ponticus) from the black sea. Vet Rec 144: 85-92.
Dailey MD, Santangelo RV, Gilmartin WG (1988). A coprological survey of helminth parasites of the Hawaiian monk seal from the Northwestern Hawaiian Islands. Marine Mammal Science 4: 125-131.
Earley EM (1975). Chromosome preparations from monolayer cell cultures. TCA Manual 1: 31-35.
Fay FH, Rausch VR, Feltz ET (1967). Cytogenetic comparison of some pinnipeds (Mammalia: Eutheria). Can J Zool 45: 773-778.
Freshney RI (ed). (1994). Culture of Animal Cells, A Manual of Basic Technique (486 pp), New York.
Fuhrman JA (1999). Marine viruses and their biogeochemical and ecological effects. Nature 399: 541-548.
Geraci JR, Aubin DJ, Barker IK, Webster RG, Hinshaw VS, Bean WJ, Ruhnke HL, Prescott JH, Early G, Baker AS, Madoff BS, Schooley RT. (1982). Mass mortality of harbor seals: Pneumonia associated with influenza A virus. Science 215: 129-131.
Gilmartin WG (1983). Recovery plan for the Hawaiian monk seal, (Monachus schauinslandi). Southwest Region, National Marine Fisheries Service, National Oceanic and Atmospheric Administration (29 pp), Long Beach, California.
Gilmartin WG, DeLong RL, Smith AW, Griner LA, Dailey MD (1980). An investigation into unusual mortality in the Hawaiian monk seal, (Monachus schauinslandi). In: Grigg RW, Pfund RT (eds), Proceedings of the Symposium on Status of Resource Investigations in the Northwestern Hawaiian Islands. University of Hawaii Sea Grant College Program (UNIHI-SEAGRANT-MR-80-04) (pp 32-41), Honolulu, Hawaii.
Gilmartin WG, Johanos TC, Gerrodette T (1987). Preliminary assessment of juvenile Hawaiian monk seal survival. Southwest Fisheries Center, National Marine Fisheries Service, Honolulu, Hawaii, H-87-16.
Harwood J (1998). What killed the monk seals? Nature 393:17-18.
Johnson AM, DeLong RL, Fiscus CH, Kenyon KW (1982). Population status of the Hawaiian monk seal (Monachus schauinslandi), 1978. J Mammalogy 63: 415-421.
Kadoi K, Mochizuki A, Ikeda T, Kamata H, Yukawa M, Inoue Y (1992). Susceptibility of a line of dolphin kidney cell culture to several herpesviruses. J Basic Microbiol 32: 227-232.
Kenyon KW, Rice DW (1959). Life history of the Hawaiian monk seal. Pacific Science 13: 215-252.
Lewis GD, Molloy SL, Greening GE, Dawson J (2000). Influence of environmental factors on virus detection by RE-PCR and cell culture. J Appl Microbiol 88: 633-640.
Liess B, Frey HR, Zaghawa A (1989). Morbillivirus in seals: Isolation and some growth characteristics in cell cultures. Dtsch Tierarztl Wochenschr 96: 180-182.
Lu Y, Aguirre AA, Braun RC, Loh PC (1998). Establishment of monk seal cell lines. In Vitro Cell Dev Biol-Animal 34: 367-369.
Lu Y, Nerurkar VR, Aguirre AA, Work TM, Balazs GH, Yanagihara R (1999). Establishment and characterization of 13 cell lines from a green turtle (Chelonia mydas) with fibropapillomas. In Vitro Cell Dev Biol 35: 389-393.
Lu Y, Tapat LM, Loh PC, Brock JA, Gose R (1995). Development of a quantal assay in primary shrimp cell culture for yellow head baculovirus (YBV) of penaeid shrimp. J Virol Methd 52: 231-236.
Mahy BWJ (1993). Seal plague virus. In: Morse SS (ed), Emerging Viruses (pp 185-193). Oxford University Press, Oxford.
Osterhaus A, Groen J, Niesters H, van de BM, Martina B, Vedder L, Vos J, Van EH, Abou-Sidi B, Barham ME (1997). Morbillivirus in monk seal mass mortality. Nature 388: 838-839.
Osterhaus A, van de BM, Vedder L, Martina B, Niesters H, Vos J, Van EH, Liem D, Baumann R, Androukaki E, Kotomatas S, Komnenou A, Abou-Sidi B, Jiddou AB, Barham ME (1998). Monk seal mortality: Virus or toxin? Vaccine 16: 979-981.
Osterhaus A, Vedder EJ (1988). Identification of virus causing recent seal deaths. Nature 335: 20-22.
Quackenbush SL, Work TM, Balazs GH, Casey RN, Rovnak J, Chaves A, duToit L, Baines JD, Parrish CR, Bowser PR, Casey JW (1998). Three closely related herpesviruses are associated with fibropapillomatosis in marine turtles. Virology 246: 392-399.
Rausch RL (1969). Diphyllobothriid cestodes from the Hawaiian monk seal, Monachus schauinslandi Matschie, from Midway Atoll. J Fish Res Board Canada 26: 947-956.
Rice DW (1960). Population dynamics of the Hawaiian monk seal. J Mammalogy 41: 376-385.
Rice DW (1977). A list of the marine mammals of the world. 3rd ed. NOAA technical report NMFS SSRF 711, Washington, DC.
Rovnak J, Quackenbush SL, Reyes RA, Baines JD, Parrish CR, Casey JW (1998). Detection of a novel bovine lymphotropic herpesvirus. J Virol 72: 4237-4242.
Smith AW, Skilling DE (1979). Viruses and virus diseases of marine mammals. J Am Vet Med Assoc 175: 918-920.
Van Bressem MF, Van Waerebeek K, Raga JA (1999). A review of virus infections of Cataceans and the potential impact of morbilliviruses, poxviruses and papillomaviruses on host population dynamics. Dis Aquat Org 38: 53-65.
VanDevanter DR, Warrener P, Bennett L, Schuttz ER, Coulter S, Garber RL, Rose TM (1996). Detection and analysis of diverse herpesviral species by consensus primer PCR. J Clin Microbiol 34: 1666-1671.
Whittow GC, Balazs GH (1979). Parasitic ulceration of the stomach in a Hawaiian monk seal (Monachus schauinslandi). 'Elepaio 38: 83-84.
Wolf K (1988). Fish viruses and fish viral diseases (p 543). Cornell Press, Ithaca and London.
Rights and permissions
About this article
Cite this article
Lu, Y., Aguirre, A.A., Hamm, C. et al. Establishment, cryopreservation, and growth of 11 cell lines prepared from a juvenile Hawaiian monk seal, Monachus schauinslandi. Methods Cell Sci 22, 115–124 (2000). https://doi.org/10.1023/A:1009816715383
Issue Date:
DOI: https://doi.org/10.1023/A:1009816715383