BeNa Culture Collection
|Culture medium||Special Medium for SF9 Cells: TNM-FH + 10% FBS|
|Subculture procedure||Recovery steps: ① The frozen vial is taken out of liquid nitrogen or -80 ℃ refrigerator and put into PE gloves, quickly submersed into a 37 ℃ water bath pan, shaken 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 5min, discard the supernatant, resuspend the cells with 1-2ml of complete media. ③The cell suspension was then added to the T25 flask containing 6-8mL of complete medium and placed in an incubator for culture.
cell subculture: ① suck the culture solution (including suspended cells) into the centrifuge tube; ② Gently blow the cell layer with PBS, blow the cell layer off and blow it away. (3) The cells obtained from the above two steps are mixed evenly and centrifuged at 1000rpm for 5min. The collected cells are divided into fresh T25 bottles according to a ratio of 1:2. Appropriate complete culture medium is added. The cell suspension is evenly mixed and cultured in an incubator. ④ 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 80%-90%.
|Growth conditions||Culture temperature 28 ℃; Air 100% in atmosphere;|
|Growth characteristics||semi-adherent and semi-suspension growth|
|Storage conditions||Liquid nitrogen|
|morphology||Epithelial cell-like, round, single cell, monolayer adherent growth, clean background|
|Separation substrate||ovaries of armyworm; spontaneous immortality; female|
|application||Sf9 cells are ovarian tissues derived from the pupae of the female Spodoptera frugiperda and can be used to replicate baculovirus expression vectors|
|Sharing mode||Public welfare sharing|
SF9 insect ovarian cells
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.
(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|
|Conclusion:||good viability, and no abnormal cell morphology, qualified|