Human small cell lung cancer cells|BNCC359344 |BNCC

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Human small cell lung cancer cells

Culture medium	RPMI-1640 complete medium: 90% RPMI-1640 + 10% FBS
Subculture procedure	Recovery steps: ① Take out the frozen vial from liquid nitrogen or -80 ℃ refrigerator and put it into PE gloves, quickly submerse into a 37 ℃ water bath, shake 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 3-5min, discard the supernatant, resuspend the cells with 1-2ml of complete media. ③The cell suspension was then added to the T25 flask containing 6-7mL of complete medium and placed in an incubator for culture. The culture flask is recommended to be cultured uprightly. cell subculture: ① centrifugation: collect cells, centrifuge at 1000rpm for 5min, 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 medium exchange method: half-volume medium exchange method can be selected; Please gently suck half of the supernatant culture medium, resuspend and mix the remaining culture medium with cell precipitation, and divide the cell suspension into a fresh T25 flask containing 8mL culture medium according to a ratio of 1:2. (3) 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 more than 2 × 106 cells/ml.
Growth conditions	Culture temperature 37 ℃; Atmosphere 5% CO2 + 95% air;
Growth characteristics	Suspension growth
Storage conditions	Liquid nitrogen
Safety level	0
morphology	Lymphoblastoid, rounded, irregular margin, many agglomerated, clean background, no pigment, no vacuoles
Sharing mode	Public welfare sharing

Human small cell lung cancer cell NCI-H69

Suspension, lymphoblast-like

No. 101668

Product format: 2ml frozen vial x 2, or centrifuge tube 15mL

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. For centrifuge tube 15mL.,please place the original centrifuge tube into incubator fir 4 hours after receiving it and then handle routinely for the cells . 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% co ₂,90% RPMI-1640 + 10% FBS. RPMI-1640:1640 culture solution, 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 7-9 days.

Subculture: gently blow the cultured suspended cells evenly, distribute them into 2-3 fresh culture liquid dishes, gently shake well and put them into an incubator for culture;

Cryopreservation: transfer the suspended cells to a centrifuge tube, centrifuge (110g,3min), resuspend the cells with 3mL frozen storage solution (50% basic medium + 40% FBS + 10% DMSO) after centrifugation, blow evenly, and divide them into 3 frozen storage tubes, use the program cooling box to freeze at -80 ℃ and transfer to liquid nitrogen storage overnight.

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 190 hours	inoculation with 20% cell solution, suspension growth rate reaches 30.0% in 18 hours, and 80.0% in 180 hours
cell morphology:	suspension, lymphoblast-like	Suspension, lymphoblast-like, round cluster growth
attached：
conclusion: good viability, no abnormal cell morphology, qualified

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