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Human brain microvascular endothelial cells & nbsp;
HBMEC
No. 350771
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 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 loss of cell viability.
Growth conditions:37 ℃,5% CO2,ECM culture solution: 500mlECM + 25mlFBS + 5mlECGS + 5mlP/S. ECM: Special Medium for ECM Endothelial Cells.
Recovery steps: (1)prepare a new 100mm culture dish containing 12ml of the above culture medium; (2)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. (3)draw the solution with a sterile pipette, drip into a new culture dish, and mix it evenly by shaking clockwise; (4) Put it into incubator (37℃,5%CO2), change the media overnight, and it will grow up in 4-6 days.
Subculture/cryopreservation: remove the medium, rinse twice with PBS , and add 2mL pancreatic enzyme substitute(TrypLE ™ Express); observe under a microscope, do not shake the culture dish during this period, when the cells detach, suck out most of the pancreatin with about 0.5ml remained, move to the incubator for digestion, and take out in about 2min. For subculture, digestion is terminated by 6mL of culture solution, and subculture at 1:2. For cryopreservation, terminate the digestion with 3mL of cryopreservation solution ( 50% base medium+40%FBS+10%DMSO ), blow evenly, dispense into 3 frozen vials, and freeze at -80 ℃ with a program cooling box. Recovery record: According to the recovery instructions, the results of the cell recovery are reported as follows: