KC-4516

CHOK1 rat CD70 Cell Line

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Background of CHOK1 rat CD70 Cell Line

CD70 (CD70 Molecule) also known as CD27L; TNFSF7, it is a Protein Coding gene. CD70 is a member of the tumor necrosis family, has been described in both hematological and solid tumors, where it increases tumor cell and regulatory T cell survival by signaling through its receptor, CD27. In solid tumors, only expression of CD70 is present on the tumor cells which can facilitate immune evasion through CD27 expression in the tumor microenvironment. Diseases associated with CD70 include Lymphoproliferative Syndrome 3 and Primary Immunodeficiency With Predisposition To Severe Viral Infection.

Specifications

Catalog Number	KC-4516
Cell Line Name	CHOK1 rat CD70 Cell Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous rat CD70 gene
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/ml
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-rat-CD70 Cell Line was generated using a lentiviral vector expressing the rat CD70 sequence.

Characterization

Figure 1: Characterization of rat CD70 overexpression in the CHOK1 rat CD70 stable clone using QPCR.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/mL Puromycin)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

Wajant H. Therapeutic targeting of CD70 and CD27. Expert Opin Ther Targets. 2016 Aug;20(8):959-73. doi: 10.1517/14728222.2016.1158812. Epub 2016 Mar 15. PMID: 26914723.
Liu K, Yang Q, Liu P, Zhu K, Zou M, Zhu Q, Yi P, Fang K, Luo Z. CD70 is a potential prognostic marker and significantly regulates cellular function in diffuse large B-cell lymphoma. PLoS One. 2024 Oct 24;19(10):e0312445. doi: 10.1371/journal.pone.0312445. PMID: 39446784; PMCID: PMC11500843..
Jacobs J, Zwaenepoel K, Rolfo C, Van den Bossche J, Deben C, Silence K, Hermans C, Smits E, Van Schil P, Lardon F, Deschoolmeester V, Pauwels P. Unlocking the potential of CD70 as a novel immunotherapeutic target for non-small cell lung cancer. Oncotarget. 2015 May 30;6(15):13462-75. doi: 10.18632/oncotarget.3880. PMID: 25951351; PMCID: PMC4537027.
Flieswasser T, Van den Eynde A, Van Audenaerde J, De Waele J, Lardon F, Riether C, de Haard H, Smits E, Pauwels P, Jacobs J. The CD70-CD27 axis in oncology: the new kids on the block. J Exp Clin Cancer Res. 2022 Jan 6;41(1):12. doi: 10.1186/s13046-021-02215-y. PMID: 34991665; PMCID: PMC8734249.