1种牙鲆囊胚期细胞系的构建及初步应用研究

doi:10.16378/j.cnki.1003-1111.24120

Abstract
Figure/Table
References
Related Citation (15)

Download: PDF (8532 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract One new embryonic multipolar morphology cell line with a clearly defined birefringent edge was established by crushing the embryos with a sterile mortar to break the membrane and carefully squeezing the embryos vertically to separate the cells as much as possible, derived from blastocysts of olive flounder Paralichthys olivaceus, designated as PoEC. The cells were cultured in DF12 complete medium, supplemented with antibiotics, fetal bovine serum (FBS), and growth factors at temperature of 25 ℃, and subcultured for 40 passages. The origin of the cell lines shwoing typical growth curve pattern, was confirmed by examining the partial sequences of the cytochrome oxidase c subunit Ⅰ (COⅠ) gene of the flounder mitochondrial DNA (mtDNA). According to the gene and activity, the cell line PoEC did not have pluripotent, whose the cell line were also successfully transfected with the green fluorescent protein (GFP) reporter gene, suggesting that it was used to investigate the gene function in the flounder or other fish. The PoEC cell line was exposed to Cr concentrations of 0, 10, 20, 30, and 40 mg/L in vitro toxicity test for 16 h, and the results showed even at concentration of 10 mg/L, Cr was toxic to the cell line in one hour, PoEC being sensitive to chromium (Cr),and used as a powerful tool for research of the toxicology of heavy metals in fish. Therefore, the cell line would be useful for biotechnological and toxicological research on marine fish as an in vitro biological system.

Key words： Paralichthys olivaceus blastocyst cell line toxicological study

Received: 12 July 2024

PACS:

S917.4

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	NIE Miaomiao
	WU Zhihao
	WANG Lijuan
	YOU Feng

Cite this article:

NIE Miaomiao,WU Zhihao,WANG Lijuan, et al. Establishment and Application of Blastocyst Cell Line of Olive Flounder Paralichthys olivaceus[J]. Fisheries Science, 2025, 44(4): 651-658.

URL:

https://www.shchkx.com/EN/10.16378/j.cnki.1003-1111.24120 OR https://www.shchkx.com/EN/Y2025/V44/I4/651

[1]BOLS N C, DAYEH V R, LEE L E J, et al. Chapter 2 use of fish cell lines in the toxicology and ecotoxicology of fish. Piscine cell lines in environmental toxicology[J]. Biochemistry and Molecular Biology of Fishes,2005,6:43-84.
[2]GOSWAMI M, YASHWANTH B S, TRUDEAU V, et al. Role and relevance of fish cell lines in advanced in vitro research[J]. Molecular Biology Reports,2022,49(3):2393-2411.
[3]THANGARAJ R S, NARENDRAKUMAR L, PRASANNAN GEETHA P, et al. Comprehensive update on inventory of finfish cell lines developed during the last decade (2010-2020)[J]. Reviews in Aquaculture,2021,13(4):2248-2288.
[4]BOLS N C. Biotechnology and aquaculture:the role of cell cultures[J]. Biotechnology Advances,1991,9(1):31-49.
[5]BAIROCH A. The cellosaurus, a cell-line knowledge resource[J]. Journal of Biomolecular Techniques,2018,29(2):25-38.
[6]LAKRA W S, RAJA SWAMINATHAN T, JOY K P. Development, characterization, conservation and storage of fish cell lines:a review[J]. Fish Physiology and Biochemistry,2011,37(1):1-20.
[7]HONG Y H, WINKLER C, SCHARTL M. Pluripotency and differentiation of embryonic stem cell lines from the medakafish (Oryzias latipes)[J]. Mechanisms of Development,1996,60(1):33-44.
[8]PENG L Y, ZHOU Y H, XU W T, et al. Generation of stable induced pluripotent stem-like cells from adult zebra fish fibroblasts[J]. International Journal of Biological Sciences,2019,15(11):2340-2349.
[9]NAKAMURA S, KOBAYASHI K, NISHIMURA T, et al. Identification of germline stem cells in the ovary of the teleost medaka[J]. Science,2010,328(5985):1561-1563.
[10]HONG Y H, LIU T M, ZHAO H B, et al. Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro[J]. Proceedings of the National Academy of Sciences of the United States of America,2004,101(21):8011-8016.
[11]GOSWAMI M, LAKRA W S, YADAV K, et al. Development of an ES-like cell culture system (RESC) from rohu, Labeo rohita (Ham. )[J]. Fish Physiology and Biochemistry,2012,38(6):1775-1783.
[12]CHEN S L, SHA Z X, YE H Q. Establishment of a pluripotent embryonic cell line from sea perch (Lateolabrax japonicus) embryos[J]. Aquaculture,2003,218(1/2/3/4):141-151.
[13]CARMEN ALVAREZ M, BÉJAR J, CHEN S L, et al. Fish ES cells and applications to biotechnology[J]. Marine Biotechnology,2007,9(2):117-127.
[14]KANG M S, OH M J, KIM Y J, et al. Establishment and characterization of two new cell lines derived from flounder, Paralichthys olivaceus (Temminck & Schlegel)[J]. Journal of Fish Diseases,2003,26(11/12):657-665.
[15]YAZAWA R, HIRONO I, OHIRA T, et al. Functional analysis of tumor necrosis factor gene promoter from Japanese flounder, Paralichthys olivaceus, using fish cell lines[J]. Developmental & Comparative Immunology,2005,29(1):73-81.
[16]ZHENG Y, PENG L M, YOU F, et al. Establishment and characterization of a fish-cell line from the brain of Japanese flounder Paralichthys olivaceus[J]. Journal of Fish Biology,2015,87(1):115-122.
[17]GAO C, SONG H Y, WANG M X, et al. Establishment and characterization of four long-term cultures of neural stem/progenitor cell lines from the Japanese flounder Paralichthys olivaceus[J]. Journal of Ocean University of China,2020,19(5):1153-1162.
[18]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.
[19]PENG L M, ZHENG Y, YOU F, et al. Establishment and characterization of a testicular sertoli cell line from olive flounder Paralichthys olivaceus[J]. Chinese Journal of Oceanology and Limnology,2016,34(5):1054-1063.
[20]NIE M M, LU Y L, ZOU C C, et al. Insight into AMPK regulation mechanism in vivo and in vitro:responses to low temperatures in the olive flounder Paralichthys olivaceus[J]. Journal of Thermal Biology,2020,91:102640.
[21]CHEN S L, REN G C, SHA Z X, et al. Establishment of a continuous embryonic cell line from Japanese flounder Paralichthys olivaceus for virus isolation[J]. Diseases of Aquatic Organisms,2004,60(3):241-246.
[22]KIM J W, OH B G, KIM J, et al. Development and characterization of a new cell line from olive flounder Paralichthys olivaceus[J]. Development & Reproduction,2018,22(3):225-234.
[23]NIE M M, ZOU C C, PENG L M, et al. Establishment and application of four long-term culture cell lines of the olive flounder Paralichthys olivaceus blastocysts[J]. Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology,2023,265:109536.
[24]NIE M M, WU Z H, YOU F. Derivation and characterization of a new embryonic cell line from the olive flounder Paralichthys olivaceus[J]. Turkish Journal of Fisheries and Aquatic Sciences,2021,21(4):159-167.
[25]COOPER J K, SYKES G, KING S, et al. Species identification in cell culture:a two-pronged molecular approach[J]. In Vitro Cellular & Developmental Biology. Animal,2007,43(10):344-351.
[26]PARODI B, ARESU O, BINI D, et al. Species identification and confirmation of human and animal cell lines:a PCR-based method[J]. BioTechniques,2002,32(2):432-434,436,438-440.
[27]WARD R D, ZEMLAK T S, INNES B H, et al. DNA barcoding Australia′s fish species[J]. Philosophical Transactions of the Royal Society of London. Series B,Biological Sciences,2005,360(1462):1847-1857.
[28]ZHENG W J, SUN L. Evaluation of housekeeping genes as references for quantitative real time RT-PCR analysis of gene expression in Japanese flounder (Paralichthys olivaceus)[J]. Fish & Shellfish Immunology,2011,30(2):638-645.
[29]SHI Z Y, CHEN X W, GU Y F. Cloning and expression pattern of alkaline phosphatase during the development of Paralichthys olivaceus[J]. Fish Physiology and Biochemistry,2011,37(3):411-424.
[30]TAJU G, ABDUL MAJEED S, NAMBI K N, et al. Application of fish cell lines for evaluating the chromium induced cytotoxicity,genotoxicity and oxidative stress[J]. Chemosphere,2017,184:1-12.
[31]BORENFREUND E, BABICH H, MARTIN-ALGUACIL N. Comparisons of two in vitro cytotoxicity assays—the neutral red (NR) and tetrazolium MTT tests[J]. Toxicology in Vitro,1988,2(1):1-6.
[32]CHEN T L, LACERTE C, WISE S S, et al. Comparative cytotoxicity and genotoxicity of particulate and soluble hexavalent chromium in human and sperm whale (Physeter macrocephalus) skin cells[J]. Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology,2012,155(1):143-150.
[33]TAJU G, ABDUL MAJEED S, NAMBI K N, et al. In vitro assay for the toxicity of silver nanoparticles using heart and gill cell lines of Catla catla and gill cell line of Labeo rohita[J]. Comparative Biochemistry and Physiology. Toxicology & Pharmacology,2014,161:41-52.
[34]ALMEIDA J L, COLE K D, PLANT A L. Standards for cell line authentication and beyond[J]. PLoS Biology,2016,14(6):e1002476.
[35]YI M S, HONG N, HONG Y H. Derivation and characterization of haploid embryonic stem cell cultures in medaka fish[J]. Nature Protocols,2010,5(8):1418-1430.
[36]LI H J, XU W T, XIANG S J, et al. Defining the pluripotent marker genes for identification of teleost fish cell pluripotency during reprogramming[J]. Frontiers in Genetics,2022,13:819682.
[37]KUMAR A, TRIPATHI G, VIMAL B, et al. Expression study of pluripotency marker genes in gold fish, Carassius auratrus[J]. International Journal of Current Microbiology and Applied Sciences,2020,9(4):639-649.
[38]HU Y C, WANG B Z, DU H J. A review on sox genes in fish[J]. Reviews in Aquaculture,2021,13(4):1986-2003.
[39]FAN Z H, LIU L Y, HUANG X H, et al. Establishment and growth responses of Nile tilapia embryonic stem-like cell lines under feeder-free condition[J]. Development,Growth & Differentiation,2017,59(2):83-93.
[40]XING J G, LEE L E J, FAN L C, et al. Initiation of a zebrafish blastula cell line on rainbow trout stromal cells and subsequent development under feeder-free conditions into a cell line,ZEB2J[J]. Zebrafish,2008,5(1):49-63.
[41]BRAMBRINK T, FOREMAN R, WELSTEAD G G, et al. Sequential expression of pluripotency markers during direct reprogramming of mouse somatic cells[J]. Cell Stem Cell,2008,2(2):151-159.
[42]MOHAMED A A, EL-HOUSEINY W, EL-MURR A E, et al. Effect of hexavalent chromium exposure on the liver and kidney tissues related to the expression of CYP450 and GST genes of Oreochromis niloticus fish:role of curcumin supplemented diet[J]. Ecotoxicology and Environmental Safety,2020,188:109890.
[43]SHAW P, MONDAL P, DEY BHOWMIK A, et al. Environmentally relevant hexavalent chromium disrupts elemental homeostasis and induces apoptosis in zebrafish liver[J]. Bulletin of Environmental Contamination and Toxicology,2022,108(4):716-724.
[44]FLEURBAIX E, PARANT M, MAUL A, et al. Toxicity of lanthanides on various fish cell lines[J]. Ecotoxicology,2022,31(7):1147-1157.
[45]WEI C Y, YANG X, KANG M J, et al. An established kidney cell line from humpback grouper (Cromileptes altivelis) and its susceptibility to bacteria and heavy metals[J]. Fish Physiology and Biochemistry,2022,48(3):521-533.