Human erythroleukemia cells|359743 |BNCC

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HEL

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 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 6-8mL of complete medium and place the flask in an incubator. Cell passage: ① Suck the culture medium (including suspended cells) into a centrifuge tube, centrifuge at 1000rpm for 5 minutes, and collect the cells; ② Gently rinse T25 bottle twice with PBS, then add 1-2mL pancreatin (0.25% Trypsin+0.02% EDTA); Observe the digestion situation under the microscope. When the cell edge shrinks and the adherent is loose (a pasteur pipette can be used to suck up some pancreatin and gently blow somewhere in the cell layer, and the cell layer can be seen to detach with naked eyes, I .e. digestion is completed, otherwise digestion is continued), directly suck out pancreatin, add 5-6mL of complete medium, gently blow the cell layer off. ③ Mix the cells obtained from the above two steps together and divide them into a fresh T25 flask at a ratio of 1:2. Add appropriate complete medium, mix the cell suspension evenly, and culture in the incubator. ④ 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 80% -90%.
Growth conditions	37°C; 5% CO₂ + 95% air
Safety level	0
morphology	Lymphoblast-like，Round，Single cells
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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