D283Med Cells
Product number:
300330
General information
Organism | Human |
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Tissue | Brain |
Disease | Medulloblastoma |
Applications | 3D cell culture, Neuroscience |
Synonyms | D283 Med, D283 MED, D283-MED, D283_Med, D-283 Med, D-283MED, D283MED, D283-Med, D-283, D283, Med 283, H283 |
Characteristics
Age | 6 years |
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Gender | Male |
Ethnicity | European |
Morphology | Epithelial |
Growth properties | Clusters in Suspension/Adherent |
Identifiers / Biosafety / Citation
Citation | D283Med (Cytion catalog number 300330) |
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Biosafety level | 1 |
Expression / Mutation
Protein expression | glutamine synthetase positive, neuron specific enolase positive, glial fibrillary acidic proteins negative, S100 (S-100) protein negative |
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Isoenzymes | AK-1, 1, ES-D, 1, G6PD, B, GLO-I, 2, Me-2, 0, PGM1, 1, PGM3, 1 |
Tumorigenic | Yes, in nude mice |
Karyotype | The karyotype is 45, xY, -7, -8, -17, -20, der(20)t(1,20)(q12,q13), 8q+, 17p+ (range = 41 to 46). This is a hypodiploid cell line with a frequency of higher ploidies of 5.4%. Three marker chromosomes are present in all cells. They are: der(20)t(1,20)(q12,q13), 8q+ and 17p+. N7, N17 and N20 have single copies. The single x is structurally normal, and the Y chromosome is present as confirmed by fluorescence microscopy. |
Handling
Culture Medium | EMEM, w: 2 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: EBSS, w: 1 mM Sodium pyruvate, w: NEAA (Cytion article number 820100c) |
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Medium supplements | Supplement the medium with 10% FBS |
Subculturing | Collect suspension cells in a 15 ml tube and carefully rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add Accutase (1-2ml per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 10 minutes, then centrifuge the cells growing in suspension and the adherent cells together. Carefully resuspend the cells and dispense into new flasks which contain fresh medium. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures | D283Med cells are shipped in a deep-frozen state on dry ice. Upon receipt, confirm that the vial remains frozen. For storage, place the cryovial immediately at temperatures below -150 degrees. If you plan to culture the cells immediately, swiftly thaw the vial by shaking it in a 37 degrees water bath with clean water and an antimicrobial agent for 40-60 seconds. Remove the vial once a small ice clump persists, ensuring it remains cold. Proceed with all subsequent steps under aseptic conditions. In a sterile flow hood, disinfect the cryovial with 70% ethanol. Then, gently open the vial and transfer the cell suspension into a 15 ml centrifuge tube pre-filled with 8 ml of room temperature culture medium. Gently mix the cells. For cell separation, centrifuge at 300 x g for 3 minutes and dispose of the supernatant. Skipping centrifugation is optional, although any residual freezing medium should be removed after 24 hours. Resuspend the pellet gently in 10 ml of fresh culture medium and divide between two T25 culture flasks. Follow the subculture protocol for subsequent steps. |
Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is rigorously excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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