KC-2778

Pan02-PDL1 Cell Line

26791

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Background of Pan02-PDL1 Cell Line

Programmed death ligand 1 (PD-L1, CD274), also known as B7-H1, is a B7-family immune checkpoint ligand expressed on antigen-presenting cells, tumor cells and activated T cells. By engaging PD-1 it suppresses effector T-cell proliferation and cytokine production and maintains peripheral tolerance. Tumor upregulation of PD-L1 drives immune escape, and anti-PD-1/PD-L1 antibodies such as atezolizumab are established cancer immunotherapies.

Specifications

Catalog Number	KC-2778
Cell Line Name	Pan02-PDL1 Cell Line
NCBI/UniProt Accession Number	NM_014143.4
Clone Number	2#
Host Cell Line	Pan02
Description	The Pan02-PDL1 cell line stably overexpresses human CD274 (PD-L1)
Quantity	One vial of frozen cells (≥2×10⁶ cells)
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+1µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split at a ratio of 1:3–1:4 every 2–3 days
Incubation	37°C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Estimated 40 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

The Pan02-PDL1 cell line was generated using a lentiviral vector expressing the human CD274 sequence.

Characterization

Figure 1: Characterization of human PDL1 overexpression in the Pan02-PDL1 stable clone using FACS.

Cell Resuscitation

Pre-warm complete culture medium (basal medium and 10% FBS) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:3–1:4 every 2–3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

Keir ME, Butte MJ, Freeman GJ, Sharpe AH. PD-1 and its ligands in tolerance and immunity. Annual Review of Immunology. 2008.
Topalian SL, Hodi FS, Brahmer JR, et al. Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. New England Journal of Medicine. 2012.
Postow MA, Chesney J, Pavlick DC, et al. Nivolumab and ipilimumab versus ipilimumab in untreated melanoma. New England Journal of Medicine. 2015.

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.