HCT-8 Cells
General information
Description | The HCT-8 line is identical to the HRT-18 cell line. The cells are positive for keratin by immunoperoxidase staining. |
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Organism | Human |
Tissue | Rectum |
Disease | Adenocarcinoma |
Synonyms | HCT 8, HCT8 |
Characteristics
Age | 67 years |
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Gender | Male |
Morphology | Epithelial-like |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | HCT-8 (Cytion catalog number 300210) |
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Biosafety level | 1 |
Expression / Mutation
Antigen expression | CDx (+/-), CDy (-), |
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Isoenzymes | AK-1, 1, ES-D, 1-2, GLO-1, 2, G6PD, B, PGM1, 1, PGM3, 1, Me-2, 1 |
Tumorigenic | In nude mice |
Viruses | Reverse Transcriptase negative |
Products | Carcinoembryonic antigen (CEA) 0.5 ng/10 exp6 cells/10 days, alkaline phosphatase, keratin |
Mutational profile | HRT-18 cells carry a mutation in codon 13 of Kras gene: GGC(Wt Gly) >GAC(Asp) |
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 |
Passaging solution | Accutase |
Doubling time | 15 hours |
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 1:4 to 1:8 is recommended |
Seeding density | 2 to 4 x 10^4 cells/cm^2 |
Fluid renewal | 2 to 3 times per week |
Freezing recovery | Fast |
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,y
CSF1PO: 12
D13S317: 8,11
D16S539: 12,13
D5S818: 13
D7S820: 10,12
TH01: 7,9.3
TPOX: 8,11
vWA: 18,19
D3S1358: 17
D21S11: 29,32.2
D18S51: 11,17
Penta E: 7,14
Penta D: 9,14
D8S1179: 15
FGA: 22
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HLA alleles |
A*: 02:01:01, 24:02:01
B*: 08:01:01, 35:01:01
C*: 04:01:01, 07:01:01
DRB1*: 03:01:01, 14:54:01
DQA1*: 01:04:01, 05:01:01
DQB1*: 02:01:01, 05:03:01
DPB1*: 01:01:01, 04:01:01
E: 01:03:02, 01:xx
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