Mouse mast cell|BNCC359406 |BNCC

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Mouse mast cell

Culture 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 ℃ 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, 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 medium exchange method: half-volume medium 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 divide 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 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	37 ℃;5% CO2 95% air;
Growth characteristics	Suspension growth
Storage conditions	Liquid nitrogen
Type	lymphoblast-like
Safety level	0
morphology	Lymphoblast-like, round, single cell, few agglomerated, clean background, no pigment, no vacuoles
Separation substrate	This is a cloned mast cell line derived from the fetal liver of (B6 XA/J)F1 mice.
Sharing mode	Public welfare sharing

MC-9 mouse mast cells

suspension, lymphoblast-like

No. : 359406

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 customer service within 24 hours. Overdue shipments are deemed to have been received well. In the form of cryopreservation tubes, they should be stored in a -80°C refrigerator in time after receipt. If they are not used for a long time, they should be transferred to liquid storage overnight. Nitrogen storage; centrifuge tube 15mL, after receipt, the centrifuge tube was placed in the original incubator for 4 hours, and then the cells were routinely operated. During resuscitation, each tube should not be kept after one use. Please operate in strict accordance with this instruction, otherwise, the reissue service will not be provided due to the inactivation of cells.

Growth conditions: 37 ℃,5% co ₂,90% RPMI-1640 + 10% FBS. RPMI-1640:1640 culture solution, containing glutamine.

Resuscitation steps:

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

(2) the frozen storage tube is taken out of liquid nitrogen or -80 ℃, bathed in water at 37 ℃ for 1~2min, and moved into the safety cabinet for resuscitation as soon as possible after complete dissolution;

(3) using a sterile straw to suck the dissolved liquid into the new plate and shake well clockwise;

(4) put in (37 ℃,5% CO₂) in the incubator, change the liquid overnight, and it will be full in 4-6 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 cryopreservation solution (50% basal medium + 40% FBS + 10% DMSO) after centrifugation, blow evenly, divide them into 3 frozen storage tubes, freeze storage at -80 ℃ with a program cooling box, and transfer to liquid nitrogen storage overnight.

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 144hours	inoculation with 20% cell solution, suspension growth rate reaches 30.0% in 18 hours, and 80.0% in 130hours
cell morphology:	suspension, lymphoblast-like	Suspension, lymphoblast-like, round
attached figure:
Conclusion:	good viability, and no abnormal cell morphology, qualified

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