Establishment, characterization and application of Largemouth bronze gudgeon (Coreius guichenoti) fin and skin cell lines in disease resistance

Abstract

Largemouth bronze gudgeon (Coreius guichenoti) is an economically important fish species in the upstream Yangtze River. However, C. guichenoti has been listed in the National Key Wild Animal List given that its wild population dramatically declined under the influence of hydroelectric projects, intensive fishing, and environmental pollution. To protect this endangered species, C. guichenoti fin (CGF) and skin (CGSK) cell lines were established in present study. Both cell types exhibited epithelial morphology, and the optimal culture conditions for CGF and CGSK were cultured at 28 °C in L-15 medium with 20% serum. Currently, CGF and CGSK have been sub-cultured up to 60 passages and 30 passages, respectively. Gene identification results of nd1, 16 s, 12 s, cytb, and coi indicated that these two cells originated from the C. guichenoti, with a chromosomal number of 2n = 50. After cryopreservation in liquid nitrogen for 6 months, cell viability remained above 70%. Subsequently, predominant expressed genes by transcriptome analysis implied that CGSK cells mainly consisted of dermis fibroblast, while CGF cell types primarily comprised myofibroblasts. The successful transfection of siRNA and pEGFP-N1 into CGF and CGSK cells were observed. Furthermore, both cell lines exhibited significant cytopathic effects (CPE) when stimulated with spring viremia of carp virus (SVCV). Therefore, the establishment of CGF and CGSK cell lines provides a solid foundation for the recovery and conservation of genetic resources, as well as immunological and toxicological studies of C. guichenoti.