Mouse myeloma cells|100204 |BNCC

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SP2/0

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 6-8mL of complete medium and place the flask in an incubator. Cell passage: ① Suck the culture medium (including suspended cells) into a centrifuge tube, centrifuge at 1000rpm for 5 minutes, and collect the cells; ② Gently rinse T25 bottle twice with PBS, then 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, 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 Mix the cells obtained from the above two steps together and divide them into a fresh T25 flask at a ratio of 1:2. Add appropriate complete medium, mix the cell suspension evenly, and culture 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，Round，Monolayer adherent growth，partially floating growth
Sharing mode	Public welfare sharing

SP2/0 mouse myeloma cells

Semi-adherent semi-suspended, lymphoblast-like

No. 100204

Culture:37 ℃,5% CO₂ CM26-1 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,CM26-1 culture solution. CM26-1 culture solution: 80% IMDM + 20% FBS. IMDM:IMDM medium, containing glutamine.

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 2-4 days.

Subculture/ cryopreservation: centrifuge the suspended cells at 110g for 3 minutes to collect the cells, rinse twice with PBS, add 2ml of 0.25%Trypsin+0.02%EDTA. Observe under the microscope, remove most of the trypsin(with 0.5ml left) when the cells detach. transfer it to the incubator for digestion about 1minutes, and take it out.

Subculture: terminate digestion of cells in culture dish with 10ml of CM26-1 culture media, re-suspend the cells in centrifuge tube with 2ml of CM26-1 culture media, mix the 12ml of the solution evenly and dispense into 3~6 culture dishes.

Cryopreservation: terminate digestion with 3ml of cryopreservation solution （90%FBS+10%DMSO), aspirate and mix evenly with the cells in centrifuge tube, 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 18 hours, the cell adherence rate ≥ 80.0% in 144 hours	adherence is observed in 18 hours, cell adherence rate ≥ 80.0% in 120 hours
cell morphology:	semi-adherent and semi-suspended, lymphoblast-like	In CM26-1 culture solution, semi-adherent and semi-suspended, lymphoblast-like, round
attached figure:
Conclusion:	good viability, no abnormal cell morphology, qualified

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