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CHO-K1
Tissue
Ovary
Synonyms
CHO K1, CHOK1, CHO cell clone K1, GM15452

Documents

CHO-K1

Cell line · Product number 603480

CHO-K1 cells are a subline derived from the CHO cell line, which was originally established in the early 1950s from a Chinese hamster ovary. CHO-K1 cells ...

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Lot
Recommended (freshest lot)
  • CoA
1

01General information

Description

CHO-K1 cells are a subline derived from the CHO cell line, which was originally established in the early 1950s from a Chinese hamster ovary. CHO-K1 cells are widely utilized in the production of therapeutic monoclonal antibodies and other biopharmaceuticals. Their extensive use in biopharmaceutical protein production and vaccines is attributed to their eukaryotic nature, which allows for proper folding, assembly, and post-translational modifications such as glycosylation, which influences the stability, efficacy, and safety of the produced proteins.

In the realm of recombinant protein production, the CHO-K1 cell line is used to express a wide array of proteins, including monoclonal antibodies, growth factors, cytokines, and enzymes. These proteins have applications in therapeutic treatments, diagnostic assays, and vaccine formulations.

CHO-K1 cells exhibit a robust growth rate and are adaptable to various culture conditions, including suspension and adherent cultures, making them highly valuable for large-scale bioproduction processes. They possess a high level of genetic stability and are used for stable cell line development as they are capable of amplifying and expressing exogenous genes efficiently, which is critical for producing high yields of recombinant proteins.

CHO-K1 chinese hamster cells can be easily transfected with a variety of vectors for gene expression, facillitating gene editing or knockdown. This flexibility allows researchers to introduce specific genes, silence genes, or even perform targeted gene editing using technologies like CRISPR-Cas9 in CHO-K1 host cells.

In conclusion, the chinese hamster CHO-K1 cells and CHO cells are pivotal in biotechnological research and biopharmaceutical production, offering a versatile platform for the study of gene function and the large-scale production of recombinant proteins.

Tissue
Ovary
Applications
This cell line is an optimal choice for toxicology, industrial biotechnology and bioproduction.
Synonyms
  • CHO K1
  • CHOK1
  • CHO cell clone K1
  • GM15452

02Classification

Organism
Chinese hamster
Sex
Female
Growth properties
Adherent
Biosafety level
1

03Characteristics

Morphology
Epithelial-like
Karyotype
Chromosome Frequency Distribution 50 Cells: 2n = 22. Stemline number is hypodiploid
Doubling time
22 hours

04Identifiers

Citation
CHO-K1 (Cytion catalog number 603480)
NCBI TaxID
10029
Cellosaurus
CVCL_0214

05Expression and susceptibility

Virus susceptibility
Vesicular stomatitis (Indiana), Getah virus Virus Resist: poliovirus 2, modoc virus, Button Willow virus
Reverse transcriptase
Negative

06Handling and culture

Culture medium
Ham's F12, w: 1.0 mM stable Glutamine, w: 1.0 mM Sodium pyruvate, w: 1.1 g/L NaHCO3 (Cytion article number 820600a)
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
1 x 104 cells/cm2 will yield in a confluent layer in about 6 days
Fluid renewal
2 to 3 times per week
Post-thaw recovery
After thawing, plate the cells at 5 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
Adult