Human colon cancer cells|BNCC338682 |BNCC

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Human colon cancer cells

Culture medium	RPMI-1640 complete medium: 90% RPMI-1640 + 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. ③The cell suspension was then added to the T25 flask containing 6-8mL of complete medium and placed in an incubator for culture. cell subculture: ① transfer the cell suspension solution into a centrifuge tube, centrifuge at 1000rpm for 5min, and 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 pasteur pipette can be used to suck up some pancreatin, and gently blow somewhere in the cell layer, , the cell layer can be seen to detach with the naked eye, that is, digestion is completed, otherwise digestion continues), 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 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	semi-adherent and semi-suspension growth
Storage conditions	Liquid nitrogen
Safety level	1
morphology	Lymphoblastic, round
Sharing mode	Public welfare sharing

COLO 205 human colon cancer cells

semi-adherent semi-suspended, epithelial cell-like

number: 338682

culture :37 ℃,5% CO₂ CM2-1 liquid media

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

biosafety level: 1, handle in ultra-clean 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.

culture conditions:37 ℃,5% CO₂,CM2-1 culture solution. CM2-1 culture solution: 90% RPMI-1640 + 10% FBS. RPMI-1640:1640 medium, containing glutamine.

Recovery steps:

(1)prepare a new 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;

(3)Put it into incubator (37℃，5%CO2）, renew the media overnight, and it will grow up in 4-6 days.

Passage / cryopreservation:The culture medium (including suspended cells) was sucked into the centrifuge tube for centrifugation (110g, 3min) and cells were collected;rinse twice with PBS, add 2ml 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 1minute, and take it out.Passage: The digestion of the above dish cells was terminated with 4mL CM2-1 culture medium, and the centrifuge tube cells were suspended with 2ml CM2-1 culture medium. A total of 6mL culture medium was sucked into the dish and mixed, and the culture was divided into 3-6 dishes.Cryopreservation:Digestion was terminated with 3mL cryopreservation solution (90%FBS+10%DMSO), then sucked out and mixed with centrifuge tube cells, divided into 3 cryopreservation tubes, which were cryopreservation at -80℃ with 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 18 hours, the cell adherence rate ≥ 80.0% in 144 hours	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 108 hours
cell morphology:	semi-adherent and semi-suspended, epithelial cell-like	in CM2-1 culture medium, semi-adherent and semi-suspended, epithelial cell-like, polygonal, round
Attached pic:
Conclusion:	good viability, no abnormal cell morphology, qualified

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