WiDr Cells
General information
Description | According to Chen TR, 1987, the WiDr cell line is a derivative of the HT-29 cell line. The cells are negative for Colon Antigen 3 expression, but positive for keratin by immunoperoxidase staining. The cells expressed p53 antigen (the p53 produced has a G -> A mutation resulting in Arg -> His at position 273). Growth of WiDr cells is inhibited by tumor necrosis factor alpha (TNF Inhibitors of dihydrofolate reductase are highly cytotoxic to WiDr cells). |
---|---|
Organism | Human |
Tissue | Colon |
Disease | Adenocarcinoma |
Synonyms | WiDR, WIDR, WiDr/S, WiDr-TC, WiDrTC, LED-WiDr, Led-WiDr |
Characteristics
Age | 44 years |
---|---|
Gender | Female |
Morphology | Epithelial-like |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | WiDr (Cytion catalog number 300377) |
---|---|
Biosafety level | 1 |
Expression / Mutation
Receptors expressed | Epidermal growth factor (EGF) |
---|---|
Protein expression | CEA positive |
Antigen expression | HLA A24, A32, B15, B18 |
Isoenzymes | PGM1, 1-2, PGM3, 1-2, G6PD, B, ES-D, 1, PEP-D, 1, 6PGD, A |
Tumorigenic | Yes, in nude mice |
Products | Carcinoembryonic antigen (CEA) 118 ng/106 cells/10 days, Colon Specific Antigen (CSAp), transforming growth factor beta, keratin |
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) |
---|---|
Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:10 to 1:20 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 |
Fluid renewal | 1 to 2 times per week |
Freezing recovery | After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
|
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. |
---|---|
STR profile |
Amelogenin: x,x
CSF1PO: 11,12
D13S317: 11
D16S539: 11,12
D5S818: 11,12
D7S820: 10
TH01: 6,9
TPOX: 8,9
vWA: 17,19
D3S1358: 15,17
D21S11: 29,30
D18S51: 13
Penta E: 14,16
Penta D: 11,13
D8S1179: 10
FGA: 20,22
|
HLA alleles |
A*: 01:01:01, 24:03:01
B*: 35:01:01, 44:03:01
C*: 04:01:01
DRB1*: 04:02:01, 07:01:01
DQA1*: 02:01:01, 03:01:01
DQB1*: 02:02:01, 03:02:01
DPB1*: 04:01:01
E: 01:01:01, 01:03:01
|