NCH612 growing culture
Referência 330121
Tamanho : 1cellcultureflask
Marca : CLS Cell Lines Service
NCH612
General information
Organism | Human |
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Tissue | Brain |
Disease | Anaplastic oligodendroglioma, WHO grade III, IDH1 mutant (R132H) |
Characteristics
Age | 39 years |
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Gender | Male |
Ethnicity | Caucasian |
Growth properties | Spheroid culture |
Identifiers / Biosafety / Citation
Citation | NCH612 (Cytion catalog number 300121) |
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Biosafety level | 1 |
Depositor | C. Herold-Mende |
Expression / Mutation
Handling
Culture Medium | DMEM:Ham's F12, w: 3.1 g/L Glucose, w: 1.6 mM L-Glutamine, w: 15 mM HEPES, w: 1.0 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a) |
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Medium supplements | Supplement the medium with 10% FBS, 5 mg/L Heparin, 20 ng/mL bFGF, 20 microgram/L EGF, 5 mg/L Insulin, 100 mg/L Transferrin, 5,2 microgram/L Na-selenit, 6,3 microgram/L Progesteron, 161,1 microgram/L Putrescin, 50 mg/L Hydrocortinson |
Subculturing | For subculturing spheroid cultures, begin by mechanically dissociating the spheroids through pipetting up and down 5 to 10 times using an Eppendorf pipette with 1000 ?l filter tips. After this, centrifuge the mixture at 300g for 5 minutes at room temperature to pellet the cells. Discard the supernatant and resuspend the cell pellet in fresh culture medium. Finally, transfer the resuspended cells into new culture vessels to promote further spheroid formation. This approach ensures efficient spheroid breakdown and readies them for continued growth in a new environment. |
Split ratio | A ratio of 1:2 to 1:5 is recommended |
Seeding density | 1 x 10^5 cells/mL |
Fluid renewal | Fresh medium must be added every 2to3 days (2to5 ml depending on the size of the cell culture flask). |
Freezing recovery | Slow. After thawing allow the cells to recover from the freezing process for at least 48 hours. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is 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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STR profile | Amelogenin: x,x CSF1PO: 11,12 D13S317: 10 D16S539: 11,13 D5S818: 11,13 D7S820: 10,11 TH01: 6,7 TPOX: 8,12 vWA: 17 D3S1358: 14,18 D21S11: 28,31 D18S51: 13 Penta E: 11,14 Penta D: 9,12 D8S1179: 13 FGA: 21 |
HLA alleles | A*: 02:01:01 B*: 57:01:01, 57:01:01G C*: 04:01:01 DRB1*: 11:01:01 DQA1*: 05:05:01 DQB1*: 03:01:01 DPB1*: 04:02:01 E: 01:03:02 |