Human chronic myeloid leukemia cells|BNCC358393 |BNCC

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Human chronic myeloid leukemia cells

Culture medium	IMDM complete medium: 90% IMDM + 10% FBS
Subculture procedure	Recovery steps: ① Take out the frozen vial from liquid nitrogen or -80 ℃ and put it into PE gloves, quickly subtract into a water bath pot at 37 ℃, shake the frozen vial to accelerate dissolution, and completely dissolve within 1 minute. (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 5min, 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. A
Growth conditions	Culture temperature 37 ℃; Atmosphere 5% CO2 + 95% air;
Growth characteristics	Suspension growth
Storage conditions	Liquid nitrogen
Type	Suspension growth
Safety level	1
morphology	Lymphoblastoid, rounded, irregular margin, single cell, clean background, no pigment, no vacuoles
application	This cell line is suitable as a transfection host. K-562 cell lines have been widely used as highly sensitive in vitro targets for natural killer assays. Cultures from the ATCC reserve have been shown to have this sensitivity for assessing natural killer activity in humans. See Pross et al. Detailed analysis for in vitro determination of NK cells, including mathematical methods for quantifying NK cell activity.
Sharing mode	Public welfare sharing

Human chronic myeloid leukemia cell K562

Suspension, lymphoblast-like

No. : 358393

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. Culture conditions: 37 ℃,5% CO₂,90% IMDM + 10% FBS.

Recovery steps:

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

② take out the frozen storage tube from liquid nitrogen or -80 ℃, take a water bath at 37 ℃ for 1~2min, and move it into the safety cabinet for recovery as soon as possible after it is completely dissolved.

③ Use a sterile straw to suck the dissolved liquid into the new plate and shake it clockwise;

④ put it into an incubator (37 ℃,5% CO₂), change the liquid overnight, and it will be 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 90hours	inoculation with 20% cell solution, suspension growth rate reaches 30.0% in 18 hours, and 80.0% in 60hours
cell morphology:	suspension, lymphoblast-like	Suspension, lymphoblast-like, round
attached figure:
conclusion: good viability, no abnormal cell morphology, qualified ;

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