Human small cell lung cancer cells|359344 |BNCC

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NCI-H69

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 3-5 minutes, discard the supernatant, and resuspend the cells in 1-2mL of complete medium. ③ Add the cell suspension into a T25 flask containing 6-7mL of complete medium and place the flask vertically in an incubator. Cell passage: ① Centrifuge method: Collect cells, centrifuge at 1000rpm for 5 minutes, discard the supernatant, add 1-2mL of culture medium and blow evenly. Divide the cell suspension into fresh T25 flasks containing 8mL of culture medium at a ratio of 1:2.② Half volume liquid exchange method: Please gently aspirate half of the supernatant culture medium, resuspend the remaining culture medium with cell sediment, and divide the cell suspension into fresh T25 flask containing 8mL of culture medium in a ratio of 1:2.③ 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 greater than 2 × 10^6 cells/mL.
Growth conditions	37°C; 5% CO₂ + 95% air
Safety level	0
morphology	Lymphoblast-like，Round，Large clusters
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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