Human cell lines
Animal cell lines
Fluorescence tagged cell lines
Media & supplements
|Growth properties||Mixed, Adherent And Suspension|
|Citation||BV2 (CLS catalog number 305156)|
|Medium supplements||10% FBS, 4 mM L-glutamine, 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 20% LADMAC conditioned media|
|Subculturing||Remove and discard culture medium. These cells grow as a mixture of floating and adherent cells. Sometimes many cells are floating, they can be harvested by centrifugation of medium instead of discarding it. Add 1.0 to 2.0 mL of Accutase solution to flask and observe cells under a microscope until the cell layer is dispersed (usually within 2 to 3 minutes). Add 4.0 to 6.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C.|
|Fluid renewal||2 - 3 times per week|
|Freeze medium||CM-1 (CLS catalog number 800150)|
|Handling of cryopreserved cultures||The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still frozen. If immediate culturing is not intended, the cryovial must be stored below -150°C after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 37 °C water bath within 40-60 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath. A small ice clump should still remain and the vial should still be cold. From now on, all operations should be carried out under aseptic conditions. Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300 x g for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this case the remains of the freeze medium have to be removed 24 hours later. Resuspend the cells carefully in 10 ml fresh cell culture medium and transfer them into two T25 cell culture flasks. All further steps are described in the Subculture section.|
|Handling of proliferating cultures||One or two cell culture flasks come filled with cell culture medium. Collect the entire medium in 1 or 2 x 50 ml centrifuge tubes, respectively. Carefully add 5 ml of cell culture medium to each T25 cell culture flask. Control the cell morphology and confluency under the microscope. Incubate at 37 °C for a minimum of 24 hours. Spin down the collected medium at 300 x g for 3 minutes to collect the cells which may have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell culture medium and transfer to a T25 cell culture flask. Incubate at 37 °C for a minimum of 24 hours.|
|Sterility||Mycoplasma contamination was excluded through PCR-based and luminescence-based mycoplasma assays. Bacterial or fungal contaminations are detected through daily visual cell monitoring.|
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