Mouse colon cancer cells|BNCC359736 |BNCC

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Mouse colon cancer cells

Culture medium	DMEM-H complete medium (including 10% FBS):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 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 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, add 5-6ml of complete medium, gently blow the cell layer, blow the cell layer off and blow it off. 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	37 ℃;5% CO2 95% air;
Growth characteristics	Adherent growth
Storage conditions	Liquid nitrogen
Type	Adherent growth
Safety level	1
morphology	Epithelial-like, polygonal
Separation substrate	Large intestine; Rectum
application	Tumorigenic in nude mice (In nude mice inoculated subcutaneously with 10 (7) cells, the tumor developed at a frequency of 100% (5/5) within 21 days)
Sharing mode	Public welfare sharing

Mouse colon cancer cell CMT93

Adherent, epithelial cell-like

No. 359736

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°C, 5% CO2, 90% DMEM-H + 10% FBS. DMEM-H: DMEM high glucose medium, containing glutamine, containing sodium pyruvate.

Recovery steps:

① Prepare a fresh 100mm culture dish containing 12ml of the above culture medium;

② 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.

③ Draw the solution with a sterile pipette, drip into a new culture dish, and mix it evenly by shaking clockwise;

④ Put it into incubator (37℃，5%CO₂）, change the media overnight, and it will grow up in 3-5 days.

Subculture/cryopreservation:remove the medium, rinse twice with PBS,and add2mL pancreatin (0.25% Trypsin + 0.02% EDTA); observe under a microscope, do not shake the culture dish during this period, when the cells detach, suck out most of the pancreatin with about 0.5ml remained, move to the incubator for digestion, and take out in about 2min. For subculture, digestion is terminated by 6mL of culture solution, and subculture at 1:2. For cryopreservation, terminate the digestion with 3mL of cryopreservation solution ( 50% base medium+40%FBS+10%DMSO ), blow evenly, dispense into 3 frozen vials, and freeze at -80 ℃ with a program cooling box,transfer to liquid nitrogen overnight for storage.

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 100 hours	adherence is observed in 18 hours, cell adherence rate ≥ 80.0% in 80 hours
cell morphology:	Adherence, epithelial cell-like	Adherent, epithelial-like, polygonal
attached:
conclusion:good viability, no abnormal cell morphology, qualified ;