Journal of Tropical Oceanography >
Establishment and characterization of gill cell line from the spotted scat Scatophagus argus
Copy editor: LIN Qiang
Received date: 2019-02-18
Request revised date: 2019-04-03
Online published: 2019-11-26
Supported by
National Natural Science Fund(41806177)
National Natural Science Fund(41741006)
Copyright
Gill is the main osmoregulatory organ in fish, and the establishment of gill cell can provide an important experimental platform to reveal the osmoregulatory mechanism. As a euryhaline fish, Scatophagus argus is an ideal animal model for such studies. This study aimed to establish the branchial cell line of S. argus. The conditions for primary culture and subculture were optimized, and cell characteristics were analyzed. The optimum growth performance was observed in the branchial cell line of S. argus at 28℃ in L-15 that contained 20% fetal bovine serum (FBS), and could be successfully subcultured in 2-4 days. The population doubling time (PDT) of SG (abbreviation for the branchial cell line established in present study) cell line was 40.8 h. Eighty-seven percent SG cells could be resuscitated after cryopreservation in liquid nitrogen. To adapt to the water environment, branchial cells could maintain homeostasis through the osmotic stress response mechanism. SG cells were grown in culture media at different osmotic pressures, and cell proliferation and morphological changes were observed in this study. SG cells could proliferate during hypotonic and hyperosmotic stress (150 and 600 mOsmol·kg -1), and the proliferation rate under hypotonic stress was 1.5 times higher than that under hyperosmotic stress. It was observed that the volume of SG cell expanded in the hypotonic medium and shrank in the hyperosmotic condition. Our results indicated that SG cells have strong osmotic tolerance, with a high adaptation in the hypoosmotic environment. The establishment of SG cell line provides basic experimental materials for the study of osmotic pressure response mechanism.
Key words: Scatophagus argus; gill cell line; primary culture; subculture
ZHOU Jianan , SU Maoliang , ZHANG Junbin . Establishment and characterization of gill cell line from the spotted scat Scatophagus argus[J]. Journal of Tropical Oceanography, 2019 , 38(6) : 90 -97 . DOI: 10.11978/2019017
表1 引物序列Tab. 1 The primer sequences |
引物名称 | 序列(5′—3′) | 产物大小/bp |
---|---|---|
18S rRNA F | GGTAACGGGGAATCAGGGT | 819 |
18S rRNA R | GTGGTGCCCTTCCGTCAA |
图7 不同渗透压刺激后的金钱鱼鳃细胞(20×)a. 低渗(150mOsmol·kg-1)刺激3h后的SG细胞; b. 等渗(300mOsmol·kg-1)培养3h的SG细胞; c.高渗(600mOsmol·kg-1)刺激3h后的SG细胞 Fig. 7 SG cells after osmotic stimulation. a. SG stimulated by hypoosmosis (150 mOsmol·kg-1) for 3h; b. SG cultured at 300 mOsmol·kg-1 for 3h; c. SG stimulated by hyperosmosis (600 mOsmol·kg-1) for 3h |
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