KC-4934

CHOK1-CD80 Cell Line

53294

Home » CHOK1-CD80 Cell Line

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	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 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.