KC-6694

Jurkat-cyno-CD8a-CD8b Cell Line

66290

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Background of Jurkat-cyno-CD8a-CD8b Cell Line

The CD8 molecule, encoded by CD8A and CD8B, forms an αβ heterodimer that serves as a key co-receptor on cytotoxic T cells, recognizing MHC class I–peptide complexes and cooperating with the TCR to initiate activation signals. Cynomolgus macaques are widely used non‑human primate models due to their close evolutionary and immunological similarity to humans, yet their CD8A/B sequences differ from human orthologs. To investigate the functional properties of cynomolgus CD8 or evaluate intervention strategies targeting these molecules in a human cellular context, it is necessary to establish systems overexpressing cynomolgus CD8A-CD8B in human cells.

Specifications

Catalog Number	KC-6694
Cell Line Name	Jurkat-cyno-CD8a-CD8b Cell Line
NCBI/UniProt Accession Number	XM_005575362.2 XM_005575360.3
Clone Number	11#
Host Cell Line	Jurkat
Description	Stable Jurkat clone expressing exogenous cynomolgus monkey CD8a and CD8b 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	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS+1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Lymphoblast
Subculture	Split saturated culture 1:4-1:6 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

Jurkat-cyno-CD8a-CD8b cell line was generated using a lentiviral vector expressing the cynomolgus monkey CD8a and CD8b sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (RPMI1640+10% FBS+1μg/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 1:4-1:6 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

Srinivasan S, Zhu C, McShan AC. Structure, function, and immunomodulation of the CD8 co-receptor. Front Immunol. 2024 Aug 26;15:1412513. doi: 10.3389/fimmu.2024.1412513. PMID: 39253084; PMCID: PMC11381289.

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.