Establishment of Cell Line of Schizothorax eurystomus Caudal Fin and Effect of Salinity and Alkalinity on Its Proliferation
DAI Jincai1, LI li1,2, LI Xuetao1, WEI Jie1, NIE Zhulan1
1. Key Laboratory of Tarim Animal Husbandry Science and Technology, Xinjiang Production and Construction Corps, College of Animal Science, Tarim University, Alar 843300, China; 2. College of Life Science, Tarim University, Alar 843300, China
Abstract：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 NaHCO3 led to increase in the proliferation of S. eurystomus caudal fin cell line at the maximal rate at NaHCO3 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.
代金彩, 李丽, 李学涛, 魏杰, 聂竹兰. 宽口裂腹鱼尾鳍细胞系的建立及其应用[J]. 水产科学, 2023, 42(1): 30-38.
DAI Jincai, LI li, LI Xuetao, WEI Jie, NIE Zhulan. Establishment of Cell Line of Schizothorax eurystomus Caudal Fin and Effect of Salinity and Alkalinity on Its Proliferation. Fisheries Science, 2023, 42(1): 30-38.
[1]王德忠.新疆的裂腹鱼亚科鱼类研究[J].干旱区研究,1998,15(4):26-32. [2]郭焱.新疆鱼类志[M].乌鲁木齐:新疆科学技术出版社,2012:34-35. [3]马燕武,郭焱,张人铭,等.新疆塔里木河水系土著鱼类区系组成与分布[J].水产学报,2009,33(6):949-956. [4]龙见全,胡仁云,邓俊强,等.野生宽口裂腹鱼的捕捉及驯化养殖技术研究[J].科学养鱼,2019(4):35-36. [5]叶尚明,李兴玖,李晓春,等.新疆塔什库尔干河水库建设对鱼类资源的影响[J].水利渔业,2004,25(4):63-65. [6]曹希全,李艳慧,魏杰,等.宽口裂腹鱼体长-体质量关系和肥满度[J].西北农业学报,2019,28(8):1380-1386. [7]魏杰,曹希全,任永丽,等.宽口裂腹鱼消化系统解剖和组织学观察[J].南方水产科学,2020,16(1):120-126. [8]魏杰,张玲,马振华,等.宽口裂腹鱼不同组织总RNA提取与质量分析[J].科技创新导报,2012,9(28):19-21. [9]WOLF K, QUIMBY M C. Established eurythermic line of fish cells in vitro[J].Science,1962,135(3508):1065-1066. [10]张念慈,杨广智.草鱼吻端组织细胞株ZC-7901及其亚株ZC-7901S1的建立和特性观察[J].水产学报,1981,5(2):111-120. [11]马辰.不同倍性泥鳅(Misgurnus anguillicaudatus)鳍细胞系的建立及特性分析[D].大连:大连海洋大学,2014. [12]李状状,李霞,王缙云,等.重金属对鱼类细胞系遗传损伤和基因表达的影响[J].水产科学,2018,37(2):278-282. [13]张晓雨,李霞,秦艳杰,等.松江鲈肾组织细胞系生物学特性研究[J].水产科学,2019,38(3):395-400. [14]王缙云,李霞,秦艳杰,等.红鳍东方鲀鳍细胞系的建立及生长特性研究[J].大连海洋大学学报,2018,33(6):703-708. [15]魏云波,刘书花,王兴民,等.石鲽鳍细胞系的建立及三丁基锡对其毒性作用的研究[J].现代农业科技,2011(19):13-14. [16]樊廷俊,耿晓芬,丛日山,等.大菱鲆鳍细胞系的建立[J].中国海洋大学学报(自然科学版),2007,37(5):759-766. [17]SWAMINATHAN T R, LAKRA W S, GOPALAK-RISHNAN A, et al. Development and characterization of a new epithelial cell line PSF from caudal fin of Green chromide, Etroplus suratensis (Bloch, 1790)[J].In Vitro Cellular & Developmental Biology-Animal,2010,46(8):647-656. [18]RESLEY M J, WEBB K A Jr, HOLT G J. Growth and survival of juvenile cobia, Rachycentron canadum, at different salinities in a recirculating aquaculture system[J].Aquaculture,2006,253(1/2/3/4):398-407. [19]YAO Z L, LAI Q F, ZHOU K, et al. Developmental biology of medaka fish (Oryzias latipes) exposed to alkalinity stress[J].Journal of Applied Ichthyology,2010,26(3):397-402. [20]赵丽慧,筴金华,张艳红,等.3种品系尼罗罗非鱼(Oreochromis niloticus)盐碱耐受性和生长比较[J].渔业科学进展,2014,35(5):26-32. [21]王梦洁,储天琪,刘峰,等.盐度胁迫对小黄鱼(Larimichthys polyactis)抗氧化酶、非特异性免疫酶和Na+/K+-ATP酶活力的影响[J].海洋学报,2021,43(2):59-66. [22]NICHOLAS HA, TERRY R, ALLEN W, et al. Evaluation of flow cytometric methods for determining population potential doubling times using cultured cells[J].Cytometry Part A,1991,12(3):234-241. [23]万颖杰,张俊磊,安静.细胞培养中细菌类微生物污染的快速检测[J].局解手术学杂志,2005,14(2):88-90. [24]金敏,黄爱华,陈照立,等.常见致病真菌通用引物PCR检测技术研究[J].环境与健康杂志,2009,26(11):969-971. [25]龚巧玲,郑龙,张焕铃,等.PCR方法检测实验动物皮肤病原真菌[J].中国比较医学杂志,2007,17(3):131-135. [26]魏云波.褐点石斑鱼(Epinephelus fuscoguttatus)三种细胞系的建立、病毒敏感性以及环境污染物对其毒性作用的研究[D].青岛:中国海洋大学,2009. [27]IMAJOH M, IKAWA T, OSHIMA S I. Characterization of a new fibroblast cell line from a tail fin of red sea bream, Pagrus major, and phylogenetic relationships of a recent RSIV isolate in Japan[J].Virus Research,2007,126(1/2):45-52. [28]WANG X L, CHEN S L, SHA Z X, et al. Establishment and characterization of a new cell line from the kidney of spotted halibut Verasper variegates[J].Journal of Ocean University of China,2010,9(2):162-168. [29]CHOI H J, LEE J H, JUNG H J, et al. Establishment and characterization of a new cell line derived from the fin of olive flounder (Paralichthys olivaceus)[J].Aquaculture,2020,528:735534. [30]ZHONG X L, RESCORLA F J. Cell surface adhesion molecules and adhesion-initiated signaling:understanding of anoikis resistance mechanisms and therapeutic opportunities[J].Cellular Signalling,2012,24(2):393-401. [31]姬广超,王晓明,苏苗苗,等.淋巴细胞的亚群分析预测其杀伤活性研究[J].临床合理用药杂志,2020,13(27):172-173. [32]徐伟,方思佳,关然,等.白术多糖对小鼠淋巴细胞的免疫调节作用[J].中国免疫学杂志,2020,36(13):1573-1577. [33]许朝花,李惠莲,岑万,等.佛罗里达文昌鱼的染色体标本制备与观察[J].福建师范大学学报(自然科学版),2020,36(4):64-69. [34]王朋朋,黄书林,张云科,等.哺乳动物细胞无血清全悬浮培养技术研究进展[J].中国畜牧兽医,2021,48(3):839-845. [35]VALE A D, COSTA-RAMOS C, SILVA D S P, et al. Cytochemical and ultrastructural study of anoikis and secondary necrosis in enterocytes detached in vivo[J].Apoptosis,2007,12(6):1069-1083. [36]刘立彭.博斯腾湖渔业及其资源增殖的意见[J].淡水渔业,1983,13(3):12-14. [37]王建,韩海东,赵求东,等.塔里木河流域水化学组成分布特征[J].干旱区研究,2013,30(1):10-15. [38]王建,韩海东,许君利,等.塔里木河流域出山径流水化学特征研究[J].中国环境科学,2021,41(4):1576-1587. [39]姚娜,宋勇,王帅,等.盐度和碱度对塔里木河叶尔羌高原鳅毒性的研究[J].西南农业学报,2018,31(2):423-428.
2023, Vol. 42 
