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Establishment,characterization and viral susceptibility of two cell lines derived from leopard wrasse Macropharyngodon geoffroy
Authors:J. Ma  S. Sun  L. Zeng  Y. Lu
Affiliation:1. Department of Public Health Sciences, John A. Burns School of Medicine, University of Hawaii at Manoa, , Honolulu, HI, 96822 U.S.A.;2. Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, , Wuhan, Hubei, China;3. College of Fisheries, Huazhong Agricultural University, , Wuhan, 430071 China
Abstract:Two new fish cell lines were established from skin (LWSK) and fin (LWFN) of leopard wrasse Macropharyngodon geoffroy. These cells grew optimally at 25° C in Leibovitz‐15 medium supplemented with 10% foetal bovine serum. Proliferation of M. geoffroy cells remained serum dependent up to cell passage 16, and cell‐plating efficiency ranged from 12 to 16%. Karyotypic analysis of these new cell lines at cell passage 8 indicated that both cell lines remained diploid with a peak chromosomal count of 144. PCR amplification of 16S mitochondrial DNA and the subsequent analysis confirmed that these cell lines were indeed derived from M. geoffroy. Results of viral challenge assays revealed that both LWSK and LWFN shared patterns of viral susceptibility similar to that of six fish viruses tested: LWSK and LWFN cells were highly permissive to channel catfish virus, spring viremia carp virus and snakehead rhabdovirus with high‐yield virus production ranging from 107·18±0·17 to 108·37±0·16 TCID50 ml?1 (mean ± s.d .). These newly established cell lines would be useful in attempts to isolate and study aquatic viruses, particularly the viral aetiology of green turtle fibropapilloma as M. geoffroy is known to be one of the common cleaner fish of green sea turtles.
Keywords:cell cryopreservation  chromosomal analysis  fish cell lines  LWFN  LWSK  primary cell culture
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