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SK-GT-4
Tissue
Esophagus, distal
Disease
adenocarcinoma
Synonyms
SKGT4, SK4

Documents

SK-GT-4

Cell line · Product number 305454

The SK-GT-4 cell line is a human esophageal adenocarcinoma model derived from primary adenocarcinoma arising in dysplastic Barrett's metaplasia of the dis...

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Lot
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  • CoA
1

01General information

Description

The SK-GT-4 cell line is a human esophageal adenocarcinoma model derived from primary adenocarcinoma arising in dysplastic Barrett's metaplasia of the distal esophagus. This cell line is part of the SK-GT series established at the Memorial Sloan-Kettering Cancer Center and has been instrumental in studying the progression of Barrett's esophagus to esophageal adenocarcinoma. SK-GT-4 cells exhibit epithelial characteristics and serve as a valuable tool for investigating molecular mechanisms of esophageal cancer, particularly those involving p53 tumor suppressor gene mutations.

Genomic studies of SK-GT-4 have identified a mutation in the TP53 gene, specifically a G to A transition at codon 175 in exon 5, resulting in an arginine-to-histidine substitution. This mutation has been correlated with chemoresistance to DNA-damaging agents such as 5-fluorouracil, mitomycin C, and cisplatin. Studies indicate that the mutational status of p53 significantly impacts the cellular response to chemotherapy, influencing cell cycle regulation and apoptosis in SK-GT-4 cells. These features make this cell line a critical model for exploring therapeutic strategies targeting p53-associated pathways.

SK-GT-4 has been utilized to evaluate tumor invasion potential and response to growth factors in esophageal adenocarcinoma. This cell line has demonstrated invasive capabilities in Boyden chamber assays, highlighting its relevance for studying metastasis. SK-GT-4 also expresses transcripts for various growth factors, including TGF-β and platelet-derived growth factor, which contribute to its aggressive phenotype. The cell line's established role in translational research underscores its utility in both basic cancer biology and drug development.

Tissue
Esophagus, distal
Disease
adenocarcinoma
Synonyms
  • SKGT4
  • SK4

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
2

03Characteristics

Morphology
Epithelial-like
Cell type
Epithelial cell
Doubling time
30-35 hours

04Identifiers

Citation
SK-GT-4 (Cytion catalog number 305454)
NCBI TaxID
9606
Cellosaurus
CVCL_2195

05Expression and susceptibility

Mutational profile
Mutation: KRAS, Simple, p.Gly12Ala (c.35G>C), Heterozygous; Mutation: PIK3CA, Simple, p.Glu542Lys (c.1624G>A), Heterozygous; Mutation: TP53, Simple, p.Gln100Ter (c.298C>T), Homozygous
Viruses
EBV -, HBV -, HCV -, HIV-1 -, HIV-2 -, HPV -, HTLV-I/-II -, MLV -

06Handling and culture

Culture medium
RPMI 1640, w: 2.1 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
Incubation atmosphere

37°C, 5% CO2, humidified atmosphere.

Supplements
Supplement the medium with 10% FBS
Dissociation reagent
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.
Seeding density
2-4*10^4/cm²
Fluid renewal
2 to 3 times per week
Thawing and culturing
  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

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.

Freeze medium
As a cryopreservation medium, we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
Storage conditions

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Shipping conditions

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

08Donor

Donor age
89 years
Ethnicity
Caucasian