WEHI-3B mouse myeloid mononuclear leukemia cells

BNCC number: 353342

Name: WEHI-3B mouse myeloid mononuclear leukemia cell

Growth characteristics: suspension growth

Growth conditions: 37 ℃,5% CO₂

Complete medium: 90% 1640 + 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. 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.

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 6-8ml complete medium and cultured in an incubator. 

Cell subculturing: 
(1 ) Centrifugation: collect cells, centrifuge at 1000rpm for 5 minutes, discard the supernatant, add 1-2mL of culture solution and blow well, and dispense the cell suspension into a fresh T25 flask containing 8ml of culture medium according to a ratio of 1:2. 

(2) Half-volume solution exchange:  the culture flask is sit for 5-10 minutes, half of the supernatant medium is gently sucked away, the remaining medium with cell precipitation is resuspended and mixed evenly, and the cell suspension is dispensed into a fresh T25 flask containing 8ml of medium according to a ratio of 1:2. 

(3) 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 subculture or cryopreservation when the cell density reaches more than 2 × 10⁶ cells/ml.

Notes:

①The cells are density-dependent, and are passaged for the first time (density reaches 80%). It is recommended to pass 1:2 (maintain cell density), and preferentially in the T25 bottle.

② if the culture bottle is sealed, put it into an incubator for cultivation after treatment, and remember to loosen the cap of the culture bottle.

③ The culture can be maintained by adding fresh medium or changing the medium. Or the culture is established by centrifugation, and then resuspended with 2X10⁵ living cells/mL. Keep the cell density between 2X10⁵ and 2X10⁶ living cells/mL. Adherent cells can be scraped by
Harvest.

Recovery record:  According to the recovery instructions, the results of the cell recovery are reported as follows:

Item	quality standard	Recovery record
viability	suspension growth rate ≥ 80.0% in 120 hours	adherence is observed in 18 hours, cell adherence rate ≥ 80.0% in 100 hours
cell morphology:	suspension, monocyte	monocytes, round
attached：
Conclusion:	good viability, and no abnormal cell morphology, qualified