Mouse microglia|BNCC337749 |BNCC

Essential Information Certificate Related Products

Mouse microglia

Culture medium	DMEM-H complete medium: 90% DMEM-H + 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. (3) The cell suspension was added to T25 flask containing 5-6mL complete medium and cultured in an incubator. cell subculture: suck out the old culture solution, rinse twice with PBS, suck the culture medium with a pasteur pipette, blow the cells off; you can also suck out the supernatant, add PBS, blow evenly, then collect the cells with a centrifuge tube, centrifuge for 3-5 minutes at 1000-1200rpm/min, resuspend and dispense into separate culture vessels.
Growth conditions	Culture temperature 37 ℃; Atmosphere 5% CO2 + 95% air;
Growth characteristics	Adherent growth
Storage conditions	Liquid nitrogen
Type	Adherent growth
Safety level	1
morphology	Epithelial cell-like, round, irregular, margin irregular, monolayer adherent growth, clean background, no pigment, no vacuoles
Sharing mode	Public welfare sharing

BV2 mouse microglia

Adherence, epithelial cell-like

No. 337749

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

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

Growth Condition:37 ℃,5% CO₂ CM1-1 culture solution

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 lose of cell viability.

Growth conditions:37 ℃,5% CO₂,CM1-1 culture solution. CM1-1 culture solution: 90% DMEM-H + 10% FBS. 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;
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.
(2)Draw the solution with a sterile pipette, drip into a new culture dish, and mix it evenly by shaking clockwise;
(3)Put it into incubator (37℃，5%CO₂）, change the media overnight, and it will grow up in 2-4 days.

Subculture/ cryopreservation:remove old medium, and rinse twice with PBS,add 12ml culture media directly,blow the cells with a pipette until they fall off completely, aspirate and dispense into 3~6 culture dishes;Cryopreservation: The suspended cells were transferred to the centrifuge tube for (110g, 3min) centrifugation，after centrifugationterminate digestion with 3ml of cryopreservation solution （90%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	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 70hours	adherence is observed in 18 hours, the cell adherence rate ≥ 80.0% in 66hours
cell morphology	adherent, epithelial cell-like	CM1-1 culture medium, adherent, epithelial cell-like, round
attached figure
conclusion	good viability, and no abnormal cell morphology, qualified