KC-2766

CHOK1-human-B7H3-IgV2-Cell-Line

26787

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Background of CHOK1-human-B7H3-IgV2-Cell-Line

B7H3, also known as CD276, is a checkpoint in the B7 family that plays an immune regulatory role in T cell responses. It is abnormally and consistently expressed in several human cancers, and its overexpression is associated with weak prognosis. B7-H3 is expressed on many cells and is a driving factor for immune evasion. B7-H3 can serve as a target for blocking monoclonal antibodies (mAbs), combination therapy, and modifying chimeric antigen receptors.

Specifications

Catalog Number	KC-2766
Cell Line Name	CHOK1-human-B7H3-IgV2-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human-B7H3-IgV2 gene
Quantity	One vial 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 1:4-1:5 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 human-B7H3-IgV2 Cell Line was generated using a lentiviral vector expressing the human B7H3-IgV2 sequence.

Characterization

Figure 1: Characterization of B7H3 overexpression in the CHOK1 human-B7H3-IgV2 stable clone using FACS.

Figure 2: Characterization of B7H3 overexpression in the CHOK1 human-B7H3-IgV2 stable clone using PCR sequence.

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 1:4-1:5 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

Mortezaee K. B7-H3 immunoregulatory roles in cancer. Biomed Pharmacother. 2023 Jul;163:114890. doi: 10.1016/j.biopha.2023.114890. Epub 2023 May 15. PMID: 37196544.
Picarda E, Ohaegbulam KC, Zang X. Molecular Pathways: Targeting B7-H3 (CD276) for Human Cancer Immunotherapy. Clin Cancer Res. 2016 Jul 15;22(14):3425-3431. doi: 10.1158/1078-0432.CCR-15-2428. Epub 2016 May 20. PMID: 27208063; PMCID: PMC4947428.
Zhao B, Li H, Xia Y, Wang Y, Wang Y, Shi Y, Xing H, Qu T, Wang Y, Ma W. Immune checkpoint of B7-H3 in cancer: from immunology to clinical immunotherapy. J Hematol Oncol. 2022 Oct 25;15(1):153. doi: 10.1186/s13045-022-01364-7. PMID: 36284349; PMCID: PMC9597993.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.