KC-4934

CHOK1-CD80 Cell Line

53294

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Background of CHOK1-CD80 Cell Line

CD80 is a co-stimulatory factor for the activation of T lymphocytes by CD86 and plays a significant role in autoimmune surveillance, humoral immune response and transplant reactions. Also known as B7, B7.1 or BB1, it has a molecular weight of 60 kD and is expressed on activated B lymphocytes, activated T lymphocytes, macrophages, peripheral blood mononuclear cells and dendritic cells. It belongs to the immunoglobulin superfamily, and its receptors are CD28 and CD152 (CTLA4). CD80 is a membrane antigen essential for the activation of T lymphocytes.

Specifications

Catalog Number	KC-4934
Cell Line Name	CHOK1-CD80 Cell Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human CD80 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 1:4-1:8 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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1 human CD80 Cell Line was generated using a lentiviral vector expressing the human CD80 sequence.

Characterization

Figure 1: Characterization of CD80 overexpression in the CHOK1 CD80 stable clone using FACS.

Figure 2: Characterization of CD80 overexpression in the CHOK1 CD80 stable clone using PCR sequencing.

Cell Resuscitation

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

Cell Freezing

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

References

1. Sansom DM, Manzotti CN, Zheng Y. What's the difference between CD80 and CD86?Trends Immunol. 2003 Jun;24(6):314-9. doi: 10.1016/s1471-4906(03)00111-x. PMID:12810107.
2.Teh YM, Lim SK, Jusoh N, Osman K, Mualif SA. CD80 Insights as Therapeutic Target in the Current and Future Treatment Options of Frequent-Relapse Minimal Change Disease. Biomed Res Int. 2021 Jan 6;2021:6671552. doi:10.1155/2021/6671552. PMID: 33506028; PMCID: PMC7806396.
3.Li L, Yang L, Jiang D. Research progress of CD80 in the development of immunotherapy drugs. Front Immunol. 2024 Nov 7;15:1496992. doi: 10.3389/fimmu.2024.1496992. Erratum in: Front Immunol. 2024 Dec 05;15:1536312. doi: 10.3389/fimmu.2024.1536312. Erratum in: Front Immunol. 2024 Dec 18;15:1538349. doi: 10.3389/fimmu.2024.1538349. PMID: 39575257; PMCID: PMC11578925.

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.