Insect ovarian cells|359598 |BNCC

Essential Information References Certificate Related Products

SF9

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	28 ℃; 100% air;
Safety level	0
morphology	Epithelial-like，Round，，Monolayer adherent growth
Sharing mode	Public welfare sharing

SF9 insect ovarian cells

semi-hanging, epithelial-like

No. 359598

Product form: 2ml frozen vial x 2, or T25 flask x 1

Safety level: 1, handle in ultra-clean table or safety cabinet

Receiving notice: if any abnormality is found on the day of receiving goods, please contact customer service within 24 hours. if it is overdue, it will be deemed as good receiving goods. The frozen storage tube shall be stored in a refrigerator at -80 ℃ after receiving the goods. If it is not used for a long time, it shall be transferred to liquid nitrogen storage overnight. The centrifuge tube is 15mL. After receiving the goods, the centrifuge tube shall be placed in the incubator for rest for 4 hours, and then the cells shall be subjected to routine operation. During recovery, each tube shall not be retained once used up. After recovery, it will be passed on to one generation and can be used normally. Please strictly follow this instruction, otherwise no reissue service will be provided if cell inactivation is caused.

Growth conditions:28 ℃,100% air, 90% TNM-FH + 10% FBS.

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 it into (28 ℃,100% air) incubator, change fluid overnight, 3-5 days full.

Passage: gently blow the cultured cells evenly, distribute them to 2~3 fresh culture liquid dishes, gently shake well and put them into incubator for culture;

Cryopreservation: gently blow the cells with the culture solution and transfer them to the centrifuge tube, centrifuge (110g,3min), resuspend the cells with 3mL of cryopreservation solution (50% basic medium + 40% FBS + 10% DMSO) after centrifugation, blow evenly, divide them into 3 cryopreservation tubes, and freeze the cells at -80 ℃ with a program cooling box, transfer overnight to liquid nitrogen for storage.

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 110hours	inoculation with 20% cell solution, suspension growth rate reaches 30.0% in 18 hours, and 80.0% in 96 hours
cell morphology:	Half-attached and half-suspended, epithelial-like	semi-hanging, epithelial cell-like, round
attached figure:
Conclusion:	good viability, and no abnormal cell morphology, qualified

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