Human colon adenocarcinoma lung metastasis cells|100320 |BNCC

Essential Information References Certificate Related Products

T84

Subculture procedure	Resuscitation steps: ① Remove the frozen vial from liquid nitrogen or -80 ℃ refrigerator and place it in PE gloves. Quickly immerse it in a 37 ℃ water bath, shake the vial to accelerate dissolution, and dissolve it completely within 1 minute; ② Add the dissolved cell solution into a centrifuge tube containing 9mL of complete medium on a ultra-clean bench, centrifuge at 1000-1200rpm for 5 minutes, discard the supernatant, and resuspend the cells in 1-2mL of complete medium. ③ Add the cell suspension into a T25 flask containing 5-6mL of complete medium and place the flask in an incubator. Cell passage: 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 Divide the cell suspension into a fresh T25 flask at a ratio of 1:2, add appropriate complete culture medium, mix the cell suspension evenly, and culture it in the incubator.; ④ Pay attention to changes in the pH value of the culture medium and cell density, change the medium regularly (2-3 times a week), and repeat the passage operation or cryopreservation when the cell density reaches 80% -90%.
Growth conditions	37°C; 5% CO₂ + 95% air
Safety level	0
morphology	Epithelial-like，Polygonal，irregular edge，Monolayer adherent growth
Sharing mode	Public welfare sharing

T84 human colon adenocarcinoma lung metastatic cells

adherent, epithelial cell-like

No. 100320

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 bacteria are well. The freeze dried culture shall be used up once and shall not be retained. The bacterial viability will resume after 1-2 generation of recovery and can be used normally. Handling instructions agar plate is enclosed. Please operate in strict accordance with this instruction, otherwise the replacement of bacteria are not be available in case of aberrant growth and loss of viability.

Growth conditions:37 ℃,5%CO₂,95% DMEM-H/F12K + 5% FBS. DMEM-H/F12K:DMEM high sugar medium and F12K medium are mixed with 1:1, containing glutamine and 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 5-6 days.

Subculture/cryopreservation: Aspirate the old culture medium, wash twice with PBS, add 2mL (/100mm dish/T25 bottle) trypsin (0.25%Trypsin+0.02%EDTA), observe under the microscope, do not shake the culture dish during this period , When the cells just fall off, aspirate most of the trypsin, leave about 0.5 mL, move it to the incubator for digestion, and take it out for about 2 minutes. Use 6 mL of culture medium to stop digestion, gently pipette evenly, and then pass 1:3; for cryopreservation, stop digestion with 3 mL of freezing medium (50% basal medium + 40% FBS + 10% DMSO), pipette evenly, Divide into 3 cryovials and freeze at -80°C in 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
Resurrection:	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 170 hours	adherence is observed in 18 hours, cell adherence rate ≥ 80.0% in 168 hours
cell morphology:	adherent, epithelial cell-like	Adherent, epithelial cell-like, polygonal growth
attached figure:
Conclusion:	good viability, and no abnormal cell morphology, qualified

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