NCI-H524 human non-small cell lung cancer cells

BNCC number: 360066

Growth properties : Suspension growth

Growth conditions: 37 ℃,5% CO₂

Complete medium: 90% RPMI-1640 + 10% FBS

Frozen storage conditions: 50% RPMI-1640 + 40% FBS + 10% DMSO

Receiving notice: if any abnormality is found on the day of receiving goods, please contact customer service within 24 hours. overdue goods will be deemed to be good! Freezing storage tube form: store in a refrigerator at -80 ℃ in time after receiving the goods. If it is not used for a long time, it should be transferred to liquid nitrogen overnight. During resuscitation, each tube shall not be retained once used up. T25 form: after receiving the goods, let it stand in the incubator for 2-3 hours, and then perform routine operation on the cells. Under aseptic conditions, transfer the original flask culture solution to a centrifuge tube, centrifuge at 1000-1200rpm/min for 5-10 min, resuspend the cells, transfer the suspended cells to a new T25 flask, add complete culture medium to 10ml, and put it into an incubator for culture. Please strictly follow this instruction, otherwise no reissue service will be provided if cell inactivation is caused.

Recovery steps:

① take out the frozen storage tube from liquid nitrogen or -80 ℃ and put it into PE gloves, quickly subtract into a 37 ℃ water bath pot, shake the frozen storage tube to accelerate dissolution, and it is advisable to dissolve it completely within 1 minute.

② add the dissolved cell fluid into a centrifuge tube filled with 9ml of complete culture medium in an ultra-clean table, centrifuge at 1000-1200rpm/min for 5 minutes, discard the supernatant, and resuspend the cells with 1-2ml of complete culture medium.

③ Then add the cell suspension to T25 bottle containing 6-8mL of complete medium, and put it into an incubator for erection culture.

Cell passage:

① centrifugation: collect cells, centrifuge at 1000rpm for 5 minutes, discard the supernatant, add 1-2mL of culture solution and blow well, and divide the cell suspension into a new T25 bottle containing 8ml of culture medium according to a ratio of 1:2.

② half-volume liquid exchange method: after the culture flask is allowed to stand 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 divided into a new T25 bottle containing 8ml of medium according to a ratio of 1:2.

③ pay attention to the change of pH value and cell density of the culture medium, and change the liquid regularly (2-3 times a week) until the cell density reaches more than 2 & times; When 10⁶/ml, repeat subculture operation or freeze storage.

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.

Recovery record: according to the recovery requirements, the above cell lines were recovered, and the recorded results were as follows:

Item	quality standard	recovery record
viability:	suspension growth rate ≥ 80.0% in 12days	suspension growth rate reaches  30.0% in 18hours, and 80.0% in 11days
cell morphology:	suspended, epithelial cell-like	suspended, epithelial cell-like, round, clustered growth
attached figure:
Conclusion:	good viability, and no abnormal cell morphology, qualified