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A427
Tissue
Lung
Disease
Carcinoma
Synonyms
A-427, A427N

Documents

A427

Cell line · Product number 300111

A427 cells originate from lung tissue, specifically a carcinoma, exhibit epithelial morphology and grow adherently. A427 cells have a doubling time of app...

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01General information

Description

A427 cells originate from lung tissue, specifically a carcinoma, exhibit epithelial morphology and grow adherently. A427 cells have a doubling time of approximately 28 hours in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS).

In ACL-3 medium, the doubling time is slightly extended to 38 hours, while in ACL-3 supplemented with bovine serum albumin (BSA), it reaches 42 hours. These variations in doubling time provide valuable insights into cell behaviour under different experimental conditions.

At passage 60, A427 cells display a hypotriploid to hypertriploid karyotype. This means the cells possess abnormal chromosomes, including dicentrics, minutes, and a large subtelocentric marker. Such karyotypic abnormalities are often associated with cancer cells and contribute to the unique characteristics of this cell line. A427 cells exhibit tumorigenic properties, allowing them to form tumours when injected into nude mice.

These tumours resemble undifferentiated adenocarcinoma, further emphasizing the relevance of this cell line in studying lung cancer and its progression. With its exceptional features, A427 cells find utility in various applications, particularly in cancer research. Their epithelial morphology and lung origin make them an ideal model for studying lung cancer and related diseases. Additionally, A427 cells are well-suited for 3D cell culture techniques, providing a more physiologically relevant environment to explore the behaviour of lung cancer cells.

Tissue
Lung
Disease
Carcinoma
Synonyms
  • A-427
  • A427N

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
Epithelial-like
Tumourigenic
Yes, in nude mice. Forms an undifferentiated tumor suggestive of adenocarcinoma.
Karyotype
P60) hypotriploid to hypertriploid with abnormalities including dicentrics, minutes and large subtelocentric marker

04Identifiers

Citation
A427 (Cytion catalog number 300111)
NCBI TaxID
9606
Cellosaurus
CVCL_1055

05Expression and susceptibility

Protein expression
P53 positive

06Handling and culture

Culture medium
EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a)
Incubation atmosphere

37°C, 5% CO2, humidified atmosphere.

Supplements
Supplement the medium with 10% FBS and 1% NEAA
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
1 x 104 cells/cm2 will result in a confluent monolayer within 3 days.
Fluid renewal
2 to 3 times per week
Post-thaw recovery
After thawing, plate the cells at 4 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
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
52 years
Ethnicity
Caucasian