Establishment of Cell Line of Schizothorax eurystomus Caudal Fin and Effect of Salinity and Alkalinity on Its Proliferation
DAI Jincai;LI li;LI Xuetao;WEI Jie;NIE Zhulan;Key Laboratory of Tarim Animal Husbandry Science and Technology, Xinjiang Production and Construction Corps,College of Animal Science, Tarim University;College of Life Science, Tarim University;
Schizostomus eurystomus is a unique species in the Tarim River system. Due to the disturbance of human activities, the change of ecological environment, and the influence of its own factors, the population of S. eurystomus has decreased sharply. The establishment of caudal fin cell lines has not been reported so far. In this study, caudal fin tissues were cultured in vitro of DME/F12 medium containing fetal bovine serum(FBS) by tissue mass method and trypsin digestion method, respectively. The caudal fin cell line was preliminarily established and the caudal fin cells were subcultured for 45 generations. The cell culture method was suspension culture, and the optimal culture medium was DME/F12. The optimal concentration of FBS was 20%. The optimum temperature was 25 ℃. Under the optimal conditions, the population doubling time of the 10th generation cells was determined. The population doubling time of S. eurystomus caudal fin cell line was 24.94 h, and the cells showed an “S” type growth. The 6th generation cells were recovered after being cryopreserved in liquid nitrogen for 180 days. By trypan blue staining,(88.72±0.99)% of S. eurystomus caudal fin cell line cells were active. S. eurystomus caudal fin cell line proliferated quickly after resuscitation and was passed normally. The sequencing results of mitochondrial 16S rRNA of the 10th generation cells were consistent with the GenBank gene sequence, and the consistency rate of S. eurystomus caudal fin cell line and NC036933.1 was 99.91%. At the molecular level, the S. eurystomus caudal fin cell line cell line was confirmed to be from S.eurystomus. NaCl salinity of 8‰ led to increase in the proliferation of S. eurystomus caudal fin cell line. The alkalinity of NaHCO_3 led to increase in the proliferation of S. eurystomus caudal fin cell line at the maximal rate at NaHCO_3 alkalinity of 7 g/L. The proliferation of S. eurystomus caudal fin cell line was increased first and then decreased with the increase in salinity.