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Weefsel
Ovary
Ziekte
Chinese hamster ovary, non-neoplastic; genetically engineered for CXCR4 surface expression (medium-high expression level)
Synoniemen
CHO-CXCR4

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CHO-CXCR4 Gemiddeld-hoog

Cell line · Productnummer 305411MH

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01Algemene informatie

Beschrijving

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The CHO-CXCR4-Medium-high cell line is a stable recombinant CHO (Chinese Hamster Ovary) cell line expressing the CXCR4 receptor at a medium-high level, approximately 9500 molecules per cell. This cell line was developed using an innovative landing pad technology, which ensures targeted integration of the CXCR4 gene at a pre-validated genomic locus. This approach results in consistent and reliable expression of the CXCR4 receptor, facilitating reproducible experimental outcomes.

CXCR4, also known as CD184, is a chemokine receptor involved in critical biological processes such as immune cell trafficking, hematopoiesis, and as a co-receptor for HIV entry into cells. The receptor's interaction with its ligand, CXCL12, is essential for the migration and homing of hematopoietic stem cells and leukocytes. In oncology, CXCR4 plays a significant role in tumor growth, metastasis, and angiogenesis, with its expression often upregulated in various cancers, including hematological malignancies. This upregulation is frequently associated with therapy resistance and poor prognosis. The expression of CXCR7 in this cell line was confirmed using flow cytometry.

Weefsel
Ovary
Ziekte
Chinese hamster ovary, non-neoplastic; genetically engineered for CXCR4 surface expression (medium-high expression level)
Toepassingen
Antibody screening; CXCR4-targeted therapy development; HIV entry research; hematopoietic stem cell biology; flow cytometry
Synoniemen
  • CHO-CXCR4

02Classificatie

Organisme
Chinese hamster
Seks
Vrouw
Groei-eigenschappen
Adhesie en suspensie
GGO-status
GMO-S1
Details over de GGO-status
GMO-S1: This CHO derivative contains a construct driving medium-to-high expression of human CXCR4 for GPCR signaling and ligand-binding analyses. This classification applies only within Germany and may differ elsewhere.
Bioveiligheidsniveau
1

03Kenmerken

Morfologie
Epithelial-like
Celtype
Epithelial cells
Verdubbelingstijd
approx. 14-16 hours

04Identificatiegegevens

Bronvermelding
CHO-CXCR4 Medium-high (Cytion catalog number 305411MH)
NCBI-belastingnummer
10029
Cellosaurus
CVCL_A8W0

05Expressie en vatbaarheid

Tot expressie gebrachte receptoren
CXCR4 (CD184)

06Behandeling en kweek

Kweekmedium

For adherent cultures: DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a)

For suspension cultures: CHO Growth Medium A (from InSCREENeX; InSCREENeX catalog number INS-ME-1039)

Incubatieomgeving

37°C, 5% CO2, humidified atmosphere.

Supplementen
For adherent cultures: Supplement the medium with 5% FBS. Add Geneticin (G418-Sulfat) to achieve a final concentration of 0.5 mg/mL.
Dissociatiereagens
For adherent cultures: Trypsin-EDTA
Heruitplanten
For routine adherent cell culture: Aspirate the old culture medium from the adherent cells, and wash them with PBS to remove any remaining medium. After aspirating the PBS, add the appropriate volume of Trypsin/EDTA solution based on the culture vessel size (e.g., 1 ml for a T25 flask, 3 ml for a T75 flask) and incubate at room temperature or 37°C for 5-10 minutes, or until the cells detach. Monitor detachment under a microscope, and gently tap the vessel if necessary to release the cells. Once detached, add complete medium to inactivate the Trypsin/EDTA, gently resuspend the cells, and transfer an aliquot of the cell suspension into a new culture vessel containing fresh medium. Place the vessel in an incubator set to 37°C with 5% CO2, and change the medium every 2-3 days.
Splitsingsverhouding
1 to 5
Zaaidichtheid
2 to 5 x 104 cells/cm2
Vloeistofverversing
2 to 3 times per week
Herstel na ontdooiing

After thawing, split the cells at a ratio of 1:2 to 1:3 in T25 flasks and allow the cells to recover from the freezing process and to adhere (for adherent cultures) for at least 24 hours.

Ontdooien en kweken
  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.

07Kwaliteitscontrole en cryopreservatie

Onvruchtbaarheid

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.

Kweekmedium invriezen
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.
Opslagvoorwaarden

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.

Verzendvoorwaarden

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.

08Donateur

Leeftijd van de donor
Adult