Human hepatic epithelial cells|BNCC354452 |BNCC

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Human hepatic epithelial cells

Culture medium	Special medium for THLE-2: 90% BEGM + 10% FBS
Subculture procedure	Recovery steps: ① Take out the frozen vial from liquid nitrogen or -80 ℃ and put it into PE gloves, quickly submerse into a 37 ℃ water bath pot, shake the frozen vial to accelerate dissolution, and it is advisable to dissolve all within 1 minute; (2) Add the dissolved cell liquid into a centrifuge tube containing 9ml of complete culture medium in an ultra-clean table, centrifuge at 1000-1200rpm/min for 5 minutes, discard the supernatant, resuspend the cells with 1-2ml of complete media. ③The cell suspension was then added to the T25 flask containing 5-6mL of complete medium and placed in an incubator for culture. Cell subculturing: ① remove the medium, rinse twice with PBS, and add 1-2mL pancreatin (0.25% Trypsin + 0.02% EDTA); (2) Observe the digestion situation under the microscope. When the cell edge shrinks and the adherent is loose (a pipette can be used to suck up a little pancreatin and gently blow somewhere in the cell layer, the cell layer can be seen to detach with naked eyes, that is, digestion is completed, otherwise digestion is continued), suck out pancreatin, add 5-6ml of complete medium, gently blow the cell layer off. ③ dispense the cell suspension into a fresh T25 flask according to a ratio of 1:2, add appropriate complete culture medium, mix the cell suspension evenly, and culture in an incubator. ④ Pay attention to the change of pH value of media and cell density, renew the media regularly (2-3 times a week), and repeat the subculture or cryopreservation when the cell density reaches 80%-90%.
Growth conditions	Culture temperature 37 ℃; Atmosphere 5% CO2 + 95% air;
Storage conditions	Liquid nitrogen
Type	Adherent growth
Safety level	2
morphology	epithelium
Separation substrate	Liver; left lobe THLE-2 ( ATCC CRL-2706 and THLE-3 ( ATCC CRL-11233) cell lines were derived from primary normal hepatocytes by infection with SV40 large T antigen. [RF84749] The virus entered the amphoteric packaging cell line PA317 by introducing the Bgl I-Hpa I fragment containing SV40 T antigen.
application	These immortalized human hepatocytes constitute an in vitro model for drug toxicology studies and studies of the etiology and pathogenesis of human hepatocellular carcinoma.
Sharing mode	Public welfare sharing