Human erythroleukocyte leukemia cell|BNCC359743 |BNCC

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Human erythroleukocyte leukemia cell

Culture medium	RPMI-1640 complete 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	Leukocyte Leukemia Cell
Safety level	1
morphology	Lymphoblast-like, round, single cell, few clubbed, clean background, no vacuoles
Sharing mode	Public welfare sharing

HEL human erythroleukocyte leukemia cell

suspension, lymphoblastoid

No. : 359743

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 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. Recovered cells in T25 culture flask, upon receipt, put the culture flask in the incubator for 4h, and then carry out handling procedure. 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 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 well clockwise;

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

Passage: 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 cryopreservation solution (50% basal medium + 40% FBS + 10% DMSO) after centrifugation, blow evenly, divide them into 3 cryopreservation tubes, freeze them at -80 ℃ with a program cooling box, and transfer them to liquid nitrogen for storage overnight.

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 90hours	inoculation with 20% cell solution, suspension growth rate reaches 30.0% in 18 hours, and 80.0% in 72hours
cell morphology:	suspension, lymphoblast-like	circular minority cell adherence
attached figure:
Conclusion:	good viability, and no abnormal cell morphology, qualified

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