Human liver cancer cells|BNCC342335 |BNCC

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Human liver cancer cells

Culture medium	DMEM-H complete medium: 85% DMEM-H + 15% 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, add 1-2mL pancreatin (0.25% Trypsin 0.02% EDTA); (2) Observe the digestion condition under the microscope, when the cell edge shrinks and the adherent is loose (use a pipette 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, I .e. digestion is completed, otherwise digestion is continued), gently blow the cell layer, blow the cell layer off and blow it away. ③ Dispense the cell suspension into a fresh T25 flask as the 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 ℃: 5% CO2 95% air in atmosphere;
Growth characteristics	Adherent growth
Storage conditions	Liquid nitrogen
Type	Adherent growth
Safety level	1
morphology	Epithelial cell-like, irregularly shaped, margin irregular, monolayer adherent growth, clean background, no pigment production, vacuoles
application	Human highly metastatic liver cancer cells, HCCLM3, have a high rate of tumor formation. The Institute of Liver Cancer, Zhongshan Hospital, Fudan University, found that MHCC97 is a highly heterogeneous cell group. Some cells have strong tumorigenicity and metastatic power, while some are weak. Therefore, different MHCC97-H and MHCC97-L of liver cancer cell lines with high and low metastasis were isolated and established. High levels of stem cell-like lateral population cells (sp) have been reported in MHCC97-H. Hepatocellular carcinoma SP cells have extremely high tumorigenicity and may be related to the metastatic potential of liver cancer. Then inoculate nude mice with human liver cancer cell line MHCC97-H, perform 3 lung metastasis screening, and take lung metastases into a liver cancer cell line MHCC97-LM3 with high spontaneous lung metastasis after subcutaneous inoculation, namely HCCLM3
Sharing mode	Public welfare sharing

HCCLM3 human liver cancer cells

Adherence, epithelial cell-like

No. 342335

Growth:37 ℃,5% CO₂CM1-1 culture solution

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

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

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-H + 10% FBS. DMEM-H:DMEM high sugar culture solution, containing glutamine and sodium pyruvate.

Recovery steps:
(1)prepare a new 100mm culture dish containing 12ml of the above culture medium;
remove the frozen vial from liquid nitrogen or refrigerator at -80℃, thaw the vial in 37°C water bath for 1-2 minutes. Transfer the vial to biosafety cabinet for culture as soon as the contents are completely thawed.
(2)draw the solution with a sterile pipette, drip into a new culture dish, and mix it evenly by shaking clockwise;
(3) Put it into incubator (37℃，5%CO2）, change the media overnight, and it will grow up in 2-4 days.

Subculture / cryopreservation: remove old medium, and rinse twice with PBS, add 6ml of 0.25%Trypsin+0.02%EDTA, do not shake the culture dish during the period. Observed under the microscope, remove most of the trypsin(with 0.5ml left) when the cells detach. transfer it to the incubator for digestion about 3minute, and take it out.

(1)Passage: terminate digestion with 12ml of CM1-1 culture media, aspirate and dispense into 3~6 culture dishes.

(2) Cryopreservation: terminate digestion with 6ml of cryopreservation solution （90%FBS+10%DMSO), aspirate and dispense into 3 frozen vials, and cryopreserve at -80 ℃ with a programmed cooling box.

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 12 hours, the cell adherence rate ≥ 80.0% in 90 hours	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 90 hours
cell morphology:	adherent, epithelial cell-like	CM1-1 culture medium, adherent, epithelial cell-like, polygonal, closely arranged
attached figure:
Conclusion:	good viability, and no abnormal cell morphology, qualified

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