Human hepatoblastoma cells|BNCC354619 |BNCC

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Human hepatoblastoma cells

Culture medium	DMEM-H complete medium: 90% DMEM-H + 10% FBS
Subculture procedure	Recovery steps: ① The frozen vial is taken out of liquid nitrogen or -80 ℃ refrigerator and put into PE gloves, quickly submersed into a 37 ℃ water bath pan, shaken The frozen vial to accelerate dissolution, and it is advisable to dissolve all within 1min; (2) Add the dissolved cell solution into a centrifuge tube containing 9ml of complete culture medium in an ultra-clean table, centrifuge at 1000-1200rpm for 5min, discard the supernatant, resuspend the cells with 1-2ml of complete media. (3) The cell suspension was added to T25 flask containing 5-6mL complete medium and cultured in an incubator. cell subculture: 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 some pancreatin and gently blow somewhere in the cell layer, and the cell layer can be seen to detach with naked eyes, I .e. digestion is completed, otherwise digestion is continued). directly 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 as 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 of medium and cell density, renew the medium 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;
Growth characteristics	Adherent growth
Storage conditions	Liquid nitrogen
Type	Adherent growth
Safety level	1
morphology	Epithelial cell-like, island-like distribution, irregular margin, clean background
Separation substrate	liver
Sharing mode	Public welfare sharing

HuH-6 human hepatoblastoma cells

adherent, epithelial cell-like

No. 354619

Product format: 2ml frozen vial x 2, or T25 flask x 1

Biosafety level: 1, handle in ultra-clear table or safety cabinet

Growth:37 ℃,5% co₂; CM1-1 culture solution

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the cells are well. For the frozen cells, it shall be stored in the refrigerator at -80 ℃ upon arrival. If they are not used for a long time, they shall be transferred to liquid nitrogen for storage overnight. Recovered cells in T25 culture flask, upon receipt, put the culture flask in the incubator for 4h, and then carry out handling procedure. During recovery, each vial shall be used up once and shall not be retained. After recovery, the cells can be passed on to the next generation and can be used normally. Please operate in strict accordance with this instruction, otherwise the replacement of cells are not be available in case of loss of cell viability.

Growth conditions:37 ℃,5% CO₂,CM1-1 culture solution. CM1-1 culture solution: 90% DMEM + 10% FBS. DMEM:DMEM culture solution, containing sodium pyruvate and glutamine. Resuscitation steps: ① 1 new 100mm plate containing 12mL of the above culture solution; (2) the frozen storage tube is taken out of liquid nitrogen or -80 ℃, bathed in water at 37 ℃ for 1~2min, and moved into the safety cabinet for resuscitation as soon as possible after complete dissolution; (3) using a sterile straw to suck the dissolved liquid into the new plate and shake well clockwise; (4) put into (37 ℃,5% co ₂) incubator, change fluid overnight, 4-6 days full.

Subculture/cryopreservation: suck out the old culture solution. after PBS cleaning twice, add 2mL(/100mm dish/T25 bottle) of pancreatin (0.25% Trypsin + 0.02% EDTA) and observe under a microscope. during this period, it is forbidden to shake the culture dish. when the cells just fall off, suck out most of the pancreatin, leave about 0.5mL, move to the incubator for digestion, and take out about 3.5min. Subculture with 6mL CM1-1 culture solution to terminate digestion, gently blow uniform cells, and then can be divided into 3~6 dishes for culture; For cryopreservation, 3mL of cryopreservation solution (90% FBS + 10% DMSO) is used to terminate digestion, blow evenly, divide into 3 frozen storage tubes, and use a program cooling box to freeze storage at -80 ℃.

Recovery record: According to the recovery instructions, the results of the cell recovery are reported as follows:

Item	quality standard	Recovery record
viability:	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 144 hours	adherence is observed in 18 hours, cell adherence rate ≥ 80.0% in 108 hours
cell morphology:	Adherence, epithelial cell-like	CM1-1 medium, adherent, epithelial cell-like, island-like distribution
attached：
Conclusion:	good viability, and no abnormal cell morphology, qualified

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