Cat.No. : CSC-RO1124 Host Cell: SHP-77
| Cat. No. | CSC-RO1124 |
| Description | This cell line is engineered to co-express human CD274 and luciferase reporter gene in SHP-77 cells. |
| Gene | CD274/Luc |
| Gene Species | Homo sapiens (Human) |
| Host Cell | SHP-77 |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Studying the interactions between immune cells and cancer cells 2. Studying the mechanisms of resistance to immune checkpoint blockade 3. High-throughput screening 4. Drug target validation |
| Quality Control | Negative for bacteria, yeast, fungi and mycoplasma. |
| Shipping | Dry ice |
| Storage | Liquid nitrogen |
| Revival | Rapidly thaw cells in a 37°C water bath. Transfer contents into a tube containing pre-warmed media. Centrifuge cells and seed into a 25 cm2 flask containing pre-warmed media. |
| Mycoplasma | Negative |
| Format | One frozen vial containing millions of cells |
| Storage | Liquid nitrogen |
| Safety Considerations |
The following safety precautions should be observed. 1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum. 2. No eating, drinking or smoking while handling the stable line. 3. Wash hands after handling the stable line and before leaving the lab. 4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells. 5. All waste should be considered hazardous. 6. Dispose of all liquid waste after each experiment and treat with bleach. |
| Ship | Dry ice |
Case Study Programmed cell death ligand 1 (PD-L1), also known as cluster of differentiation (CD274) or B7 homolog 1 (B7-H1), is a type 1 transmembrane protein involved in regulating cellular and humoral immune responses. The interaction of PD-L1 (CD274) with its receptor PD-1 provides stimulatory and inhibitory signals that regulate T cell activation and tolerance during pregnancy, tissue allograft transplantation, autoimmune diseases, and malignant transformation. The aim of this study was to elucidate the role of PD-L1 (CD274) in peritoneal dissemination. The study showed that mouse ovarian cancer cells induced PD-L1 (CD274) expression when they encountered lymphocytes during peritoneal dissemination in vivo and when co-cultured with lymphocytes in vitro. When PD-L1 was overexpressed, tumor cell lysis by CTLs was attenuated, whereas when it was silenced, tumor cell lysis by CTLs was enhanced. PD-L1 (CD274) overexpression inhibited CTL aggregation and degranulation. The CTL gene expression profile caused by PD-L1 (CD274) overexpressing ovarian cancer was associated with CTL exhaustion. In mouse models, PD-L1 depletion resulted in suppressed peritoneal tumor growth and prolonged survival.
To investigate the effects of PD-L1 (CD274) expression on tumor cells, the researchers established PD-L1 (CD274)-overexpressing cell lines (denoted as ID8-pdl1 and HM1-pdl1) and PD-L1 (CD274)-deficient cell lines (denoted as ID8-Mirpdl1 and HM1-Mirpdl1) from ID8 and HM-1. Cell proliferation assays showed that the proliferation curves of PD-L1 (CD274)-manipulated cell lines were similar to those of control cell lines (Figure 1A), indicating that PD-L1 (CD274) expression did not affect cell proliferation in vitro. Cytotoxicity assays showed that high levels of target cell lysis were observed in ID8-Mirpdl1 cells, while low levels of target cell lysis were observed in PD-L1 (CD274)-overexpressing ID8 cells (Figure 2B), indicating that antigen-specific cell lysis by CTLs is inhibited by PD-L1 and can be promoted by PD-L1 depletion. In addition, they evaluated changes in CTLs after encountering tumor-associated PD-L1 (CD274). CTLs lyse target cells by secreting perforin and granzymes, and CD107a is a surface marker of activated CTL degranulation. CD107a expression in CTLs co-cultured with ID8-pdl1 was weaker than that in controls, indicating that T cell degranulation after antigen stimulation was inhibited by tumor-associated PD-L1 (CD274) (Figure 1C).
Figure 1. PD-L1 protects tumor cells from CTL attack. A, Cell proliferation assay of PD-L1-manipulated HM-1 cell line (left) and ID8 cell line (right). B, Cytotoxicity assay of PD-L1-manipulated ID8 cell line. C, CD107a+ CTLs after co-incubation with OVA-loaded ID8-Mirpdl1, OVA-loaded ID8-control, OVA-loaded ID8-pdl1, or ID8-control without OVA. D, Microscope images of activated GFP+ CTLs after 136 minutes of co-incubation with ID8-control (left) or ID8-pdl1 (right). (Abiko K, et al. 2013)
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As a researcher focused on immunotherapy, reproducibility is key. The CD274/Luc Stable Cell Line - SHP-77 has provided us with highly reproducible results, enabling us to confidently advance our studies on PD-L1 inhibitors.
The cell line is incredibly easy to culture and handle. The consistent growth rates and straightforward maintenance protocols mean even junior lab members can manage it with confidence, reducing the learning curve and minimizing potential errors.
With the luciferase reporter built into the CD274 expression system, we can easily and accurately measure activity without additional constructs or convoluted procedures.
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