C3H/10T1/2, Clone 8
|Description||The cells are very sensitive to post confluence inhibition of cell division, do not produce tumors in syngeneic mice, have no background of spontaneous transformation, nor do they contain overt endogenous transforming murine leukemia or sarcoma viruses.The cells are contact sensitive.There is no detectable background spontaneous transformation.They are highly susceptible to transformation by chemical agents.Tested and found negative for ectromelia virus(mousepox).Note: the inoculation density, feeding and harvesting schedules must be followed rigidly if the line is to retain its essential characteristics.The batch of serum used for growth and for transformation assays may affect both the morphology of this line and the results obtained.Monolayers established and maintained for the standard transformation assay should be free of all foci after 6 weeks.The depositor recommends that the line be used between the 5th and 15th passages only.|
|Synonyms||C3H/10T1/2 Clone 8, C3H/10T1/2-clone8, C3H/10T1/2 CL8, C3H10T1/2 clone8, C3H10T1/2CL8, 10T1/2(clone8), 10T1/2, C3H10T1-2, C3H10T1/2, C3H-10T1/2, C3H 10T1/2, C3H/10T1/2|
Identifiers / Biosafety / Citation
|Citation||C3H/10T1/2, Clone 8 (CLS catalog number 305164)|
Expression / Mutation / Susceptibility
|Medium supplements||10% FBS, 2 mM L-glutamine|
|Subculturing||Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10 ml for T75 cell culture flasks). Add Accutase (1-2 ml per T25, 2.5 ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300 g, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium.|
|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.|
Quality control / Genetic profile / HLA
|Sterility||Mycoplasma contamination was excluded through PCR-based and luminescence-based mycoplasma assays. Bacterial or fungal contaminations are detected through daily visual cell monitoring.|
References "C3H/10T1/2, Clone 8 "
To identify mycoplasma contaminations we perform PCR-based and luminescence-based mycoplasma assays. We further determine any bacterial or fungal contamination through our standardized manufacturing processes.
Besides genomic DNA, RNA, cell pellets, and cell lysates in bulk, we can offer large quantities of assay-ready cells, plated cells in multiple formats, and frozen or growing cells. Contact us to receive a quote.
Each manufactured batch of cell lines* is authenticated via STR analysis. Contact us if you require a publication-ready STR report for your cells (*human, hamster, mouse, rat, and dog cells).
HLA characterization is available from more than 200 cell lines. HLA class I -A, B, C, and Class II HLA-DPA1, -DPB1, -DQA1, DQB1, and DRB1 alleles were obtained by next-generation sequencing methodologies (NGS) for class I and class II alleles.