|Applications||3D cell culture, Immuno-oncology|
|Synonyms||Du4475, DU-4475, Du-4475, DU 4475, Du 4475, Duke University 4475|
|Growth properties||Clusters in Suspension|
Identifiers / Biosafety / Citation
|Citation||DU4475 (CLS catalog number 300371)|
Expression / Mutation / Susceptibility
|Isoenzymes||AK-1, 1, ES-D, 1, G6PD, B, GLO-I, 2, Me-2, 2, PGM1, 1-2, PGM3, 1|
|Tumorigenic||Yes, in nude mice|
|Viruses||EBV -, HBV -, HCV -, HIV-1 -, HIV-2 -, HTLV-1/2 -, MLV -, SMRV -|
|Karyotype||human flat-moded near-tetraploid karyotype with 12% polyploidy - 88-93XXXX, +1, +1, -5, -6, +9, -10, -10, +15, +15, -16, -16, +22, +4mar, i(1q)x2, ?add(1)(p35-36)x2, ?i(5p)x2, add(6)(p11), add(6)(p1?), del(6)(q25), add(9)(q35), del(11)(q24)x2, add(15)(p11)x2, add(17)(p1?)x2, del(21)(q22.2)x2 - sideline with -20, -20, +del(7)(p11) - gain of 1q and loss of 6q typical in breast carcinoma - resembles published karyotype|
|Culture Medium||RPMI 1640|
|Medium supplements||10% FBS|
|Passaging solution||The utilization of a passaging solution is not necessary when passaging cells that are cultured in suspension. The appropriate procedure is to dilute the cells in accordance with the indicated guidelines.|
|Subculturing||Cultures can be maintained by addition or replacement of fresh medium. Start cultures at 2 x 10^5 cells/ml and maintain between 1x10^5 and 1x10^6 cells/ml.|
|Freeze medium||CM-1 (CLS catalog number 800100) or CM-ACF (CLS catalog number 806100)|
|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 degree Celsius after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 37 degree Celsius 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 degree Celsius 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 degree Celsius for a minimum of 24 hours.|
Quality control / Genetic profile / HLA
Penta E: 7,13
Penta D: 13,14
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.