Human cell lines
Animal cell lines
Fluorescence tagged cell lines
Media & supplements
|Description||This is a 6-thioguanine resistant variant of CCRF-CEM.It is deficient in Hypoxanthine phosphoribosyl transferase(HPRT, HGPRT) and secretes high titers of an immunosuppressive factor.This line may be used as a fusion partner for making human T cell hybridomas.suspensionimmunosuppressive factorCultures can be maintained by addition or replacement of fresh medium. Start cultures at 2×10 exp5 cells/ml and maintain between 1×10 exp5 and 1×10 exp6 cells/ml.Handle as potentially biohazardous material under at least Biosafety Level 2 containment.|
|Disease||T-cell acute lymphoblastic leukemia|
|Citation||6T-CEM (CLS catalog number 305132)|
|Medium supplements||10% FBS|
|Subculturing||Resuspend cell suspension in the flask and take representative aliquote to count the cell number per ml. Dilute cell suspension to 1 x 10^5 cells/ml with fresh medium and transfer into new flasks.|
|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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