BeNa Culture Collection
|Culture medium||RPMI-1640 complete medium: 80% RPMI-1640 + 20% FBS|
|Subculture procedure||Recovery steps: ① Take out the frozen vial from liquid nitrogen or -80 ℃ refrigerator and put it into PE gloves, quickly submerse into a 37 ℃ water bath, shake the frozen vial to accelerate dissolution, and it is advisable to dissolve all within 1min; (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 3-5min, discard the supernatant, resuspend the cells with 1-2ml of complete media. ③The cell suspension was then added to the T25 flask containing 6-7mL of complete medium and placed in an incubator for culture. The culture flask is recommended to be cultured uprightly.
cell subculture: ① centrifugation: collect cells, centrifuge at 1000rpm for 5min, discard the supernatant, resuspend the cells with 1-2ml of complete medium, 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 solution exchange method: half-volume solution exchange method can be selected; Please gently suck half of the supernatant culture medium, resuspend and mix the remaining culture medium with cell precipitation, and dispense the cell suspension into a fresh T25 flask containing 8mL culture medium according to a ratio of 1:2. (3) Pay attention to the change of pH value of media and cell density, renew the media 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||B cell lymphoma|
|morphology||Lymphoblast-like, round, multiple clusters, black spots in cell secretion|
|Separation substrate||bone marrow|
|Sharing mode||Public welfare sharing|
2. No.: 338435
3. Growth properties : □ wall ■ suspension □ semi-suspension and semi-wall
4. Growth conditions:
|culture medium||90% IMDM|
|air condition||5% CO2,95% AIR|
|frozen storage conditions||culture medium 50%, serum 40%, DMSO 10%|
|a bottle of cells||T25|
|cell culture and operation instructions||1 copy|
1 Upon arrival, it is suggested to sit the cells in a incubator for about 4 hours, and then renew the media for recovery or subculture according to the cell density.
2 Collect the cells by centrifuge, re-suspend the collected cells with 10ml of complete media, transfer to fresh culture flask/vessel and cultivate overnight, and dispense into separate vials for subculture according to the cell density.
3 Clustered growth: agitate the culture flask to disperse the clustered cells and continue to recover or subculture.
Recovery and subculture procedure of cells ( under strict aseptic conditions )
1 The suspension cells is normally handled by changing half of the solution and sub-cultured in separate flasks, that is, transfer half of the suspension liquid to a fresh flask/vessel, add appropriate complete medium and culture it in the incubator; It can also be sub-cultured in separate flasks according to cell density.
2 Pay attention to the change of pH value of medium and cell density, renew the medium regularly, and repeat the subculture or cryopreservation when the cell density reaches 70%-80%.
Special attention: (if handle in public laboratory or first time to culture the cells, it is recommended to add penicillin streptomycin into the media)
1 Upon the receipt of the cells, renew the solution with fresh 10% FBS medium as soon as possible. It is not recommended to use the medium used for transportation in the original flask.
2 Please take photos and contact the technicians in time if the culture flask leaks upon the receipt of cells.
3 Any compliant on the cells, please take photos and contctat our technicians.