Mouse lung cancer cells|BNCC338394 |BNCC

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Mouse lung cancer cells

Culture medium	DMEM-H complete medium: 90% DMEM-H + 10% FBS
Subculture procedure	Recovery steps: ① Take out the frozen vial from liquid nitrogen or -80 ℃ and put it into PE gloves, quickly subtract into a water bath pot at 37 ℃, shake the frozen vial to accelerate dissolution, and it is advisable to dissolve it completely within 1 minute; (2) Add the dissolved cell solution to 9ml in the ultra-clean table; In the centrifuge tube of complete culture medium, centrifuge at 1000-1200rpm/min for 3-5 minutes, discard the supernatant, resuspend the cells with 1-2ml of complete media . ③The cell suspension was then added to the T25 flask containing 6-8mL of complete medium and placed in an incubator for culture. Cell subculturing: ① transfer the cell suspension solution into a centrifuge tube, centrifuge for 5 minutes at 1000RPM to collect cells; (2) rinse T25 flask twice with PBS, add 1-2mL pancreatin (0.25% Trypsin + 0.02% EDTA); 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 on the cell layer, the cell layer can be seen to detach with the naked eye, that is, digestion is completed, otherwise digestion is continued), directly suck out pancreatin, add 5-6ml of complete medium, gently blow the cell layer off. (3) The cells obtained from the above two steps are evenly mixed and then dispensed into fresh T25 flasks as a ratio of 1:2. add appropriate complete medium, mix the cell suspension evenly, and culture in an incubator. ④ Pay attention to the change of PH value and cell density of the culture medium, change the liquid regularly (2-3 times a week), and repeat the passage operation or cryopreservation when the cell density reaches 80%-90%.
Growth conditions	37 ℃,5% CO2
Growth characteristics	Half paste and half suspension
Storage conditions	Liquid nitrogen
Safety level	1
Separation substrate	Lung
application	The line is widely used as a model of metastasis and can be used to study the mechanism of cancer chemotherapeutic agents.
Sharing mode	Public welfare sharing

LL/2(LLC1) mouse lung cancer cells

BNCC number : 338394

Growth characteristics: Semi-suspended semi-adherent growth

Growthconditions : 37 ℃,5% CO2

Complete medium : 90% DMEM-H + 10% FBS

Cryopreservation conditions : 50% basal medium + 40% FBS + 10% DMSO

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 2-3h, and then carry out handling procedure. Under sterile conditions, transfer the culture solution in T25 flask to a centrifuge tube, centrifuge at 1000-1200rpm/min for 5 ~ 10 minutes, resuspend the cells, transfer the suspended cells to a fresh T25 flask, add 10ml of complete medium, and put them into the incubator.Please operate in strict accordance with this instruction, otherwise the replacement of cells are not be available in case of loss of cell viability.

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, shaken the frozen vial to accelerate dissolution, and it is advisable to dissolve all within 1min;
(2)Put 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.
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 pasteur 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 the ratio of 1:2, add appropriate complete culture medium, make the cell suspension well distributed, 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 passage operation or cryopreservation when the cell density reaches 80%-90%.
Notes:
(1)The cells are density dependent. Initiate the subcultivation in T25 flask when the cell density reaches 80%; A subcultivation ratio of 1:2 is recommended.
(2)If the cells in sealed culture bottle, put it into the incubator for cultivation after handling, with the cap unscrewed.
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 80hours	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 72hours
cell morphology:	fibroblast-like	fibroblast-like, short spindle-shaped, round
attached figure:
Conclusion:	good viability, and no abnormal cell morphology, qualified