BeNa Culture Collection
info@bncc.com
| 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,Single cells |
| Sharing mode | Public welfare sharing |
EL4 mouse lymphoma cell
suspended, lymphoblastic
Number: 356316
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% CO2,90% DMEM-H + 10% HS. DMEM-H:DMEM high sugar culture solution, containing glutamine and sodium pyruvate.
Recovery steps:
(1)prepare a new 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 2-4 days.
Subculture:the suspended cells were gently beaten and evenly distributed into 1-3 fresh culture medium dishes. After being shaken well, the cells were put into the incubator for culture
Cryopreservation: The suspended cells were transferred to the centrifuge tube for (110g, 3min) centrifugation,after centrifugationterminate digestion with 3ml of cryopreservation solution (50% base medium+40%FBS+10%DMSO), aspirate and dispense into 3 frozen vials, and cryopreserve at -80 ℃ with a programmed 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 72hours | inoculation with 20% cell solution, suspension growth rate reaches 30.0% in 18hours, and 80.0% in 66hours |
| cell morphology: | suspended, lymphoblast-like | suspended, lymphoblast-like, round |
| attached figure: | ![]() |
![]() |
| conclusion: | good viability, and no abnormal cell morphology, qualified | |
Super Tube(Super Tube)
100297
Y79(Y79)
100707
bEnd.3[BEND3](bEnd.3[BEND3])
337672
MC38(MC38)
337716
MH7A(MH7A)
337864
MADB106(MADB106)
339574