Human breast cancer epithelial cells|BNCC359451 |BNCC

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Human breast cancer epithelial 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 ℃ and put it into PE gloves, quickly submerse into a 37 ℃ water bath pot, shake the frozen vial to accelerate dissolution, and it is advisable to dissolve all within 1 minute; (2) Add the dissolved cell liquid into a centrifuge tube containing 9ml of complete culture medium in an ultra-clean table, centrifuge at 1000-1200rpm/min for 5 minutes, discard the supernatant, resuspend the cells with 1-2ml of complete media. ③The cell suspension was then added to a T25 flask containing 6-8mL complete medium and placed in an incubator for upright cultivation. 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 liquid medium exchange: 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 fresh T25 flask containing 8ml of 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
Type	Breast cancer epithelial cells
Safety level	1
morphology	Epithelial cell-like, round, oblong
Separation substrate	Chest; breast
Sharing mode	Public welfare sharing

DU4475 human breast cancer epithelial cells

suspension, epithelial cells & nbsp; sample

No. 359451

Product format: 2ml frozen vial x 2, or T25 flask x 1

Biosafety level: 1, handle in ultra-clean table or safety cabinet

Receiving notice: if any abnormality is found on the day of receiving goods, please contact customer service within 24 hours. if it is overdue, it will be deemed as good receiving goods. The frozen storage tube shall be stored in a refrigerator at -80 ℃ after receiving the goods. If it is not used for a long time, it shall be transferred to liquid nitrogen storage overnight. The centrifuge tube is 15mL. After receiving the goods, the centrifuge tube shall be placed in the incubator for rest for 4 hours, and then the cells shall be subjected to routine operation. During recovery, each tube shall not be retained once used up. After recovery, it will be passed on to one generation and can be used normally. Please strictly follow this instruction, otherwise no reissue service will be provided if cell inactivation is caused.

Growth conditions:37 ℃,5% co ₂,90% RPMI-1640 + 10% FBS. RPMI-1640:1640 culture solution, containing glutamine.

Resuscitation steps:

(1) 1 new 100mm plate containing 12mL of the above culture solution;

(2) the frozen storage tube is taken out of liquid nitrogen or -80 ℃, bathed in water at 37 ℃ for 1~2min, and moved into the safety cabinet for resuscitation as soon as possible after complete dissolution;

(3) using a sterile straw to suck the dissolved liquid into the new plate and shake well clockwise;

(4) put in (37 ℃,5% CO₂) in the incubator, change the liquid overnight and full in 2-4 days.

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

Cryopreservation : transfer the suspended cells to a centrifuge tube, centrifuge (110g,3min), resuspend the cells with 3mL of frozen storage solution (50% basic medium + 40% FBS + 10% DMSO) after centrifugation, blow evenly, divide them into 3 frozen storage tubes, and freeze storage at -80 ℃ with a program 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:	suspension growth rate ≥ 80.0% in 96hours	inoculation with 20% cell solution, suspension growth rate reaches 30.0% in 18 hours, and 80.0% in 90hours
cell morphology:	fibroblast-like	Fibroblast-like, long spindle-shaped
attached figure:
Conclusion:	good viability, and no abnormal cell morphology, qualified

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