Cat.No. : CSC-CAR0002 Host Cell: THP-1
| Cat. No. | CSC-CAR0002 |
| Description | This cell line is a monoclonal cell line that is engineered to stably overexpress chimeric antigen receptor (CAR) targeting human CD20 (anti-CD20 CAR). |
| Gene | CD20 CAR |
| Host Cell | THP-1 |
| Host Cell Species | Homo sapiens (Human) |
| Stability | Validated for at least 10 passages |
| Application | 1. Gene expression studies 2. Signaling pathway research 3. Drug screening and toxicology 4. Disease research |
| 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 |
CAR-T (chimeric antigen receptor-engineered T) cell therapy is a promising treatment option for hematological malignancies, particularly lymphoma and B-cell leukemia. The goal of the research was to develop an Anti-hCD20-CAR(3G) Stable Cell Line to improve CAR-T cell therapy for hematological malignancies, particularly lymphoma and B-cell leukemia. This advancement addresses the tumor-associated antigen heterogeneity and T-cell exhaustion that limit the effectiveness of CAR-T therapy in solid tumors. In order to reduce PD-1-dependent inhibitory signals, they modified CAR-T cells to produce an anti-PD-1 single-chain variable fragment (scFv). The process entailed producing anti-PD-1 scFv-secreting CAR-T cells, which lessen the inhibitory effects on neighboring tumor-specific non-CAR-T cells as well as CAR-T cells themselves.
Figure 1. For two days, they cocultured 3LL-hCD20 cells at different effector-to-target ratios with anti-hCD20 conventional CAR-T, anti-hCD20 scFv CAR-T, and activated T cells without gene transfection. The research verified that, in comparison to conventional CAR-T cells, scFv CAR-T cells exhibited elevated cytotoxic activity and cell count, and generated a greater amount of IFN-γ, particularly in environments that replicated a high tumor cell burden. (Nakajima M, et al., 2019)
A: Surface expression detection: Use anti-CD20 antibody labeling through flow cytometry to detect the expression of CAR on the cell surface. This method can quantitatively analyze the expression level of CAR on the cell surface. Protein detection: Use Western blot technology to detect the presence and expression level of CAR protein to ensure the stable expression of CAR in cells. Gene expression analysis: qPCR was used to detect the mRNA level of CAR to confirm the expression of the gene and further verify the stability of CAR at the transcription level.
A: Co-culture experiment: CART cells are co-cultured with CD20-expressing target cells (such as Raji cells), and control cells (such as K562 cells) that do not express CD20 are set to evaluate the specific killing activity of CART cells. Cell viability assay: Evaluate the death rate of target cells through cytotoxicity analysis (such as LDH release assay or Annexin V/PI staining in flow cytometry) to confirm the killing effect of CART cells. Cytokine release analysis: detect the secretion of key cytokines (such as IFNγ, IL2) in the co-culture system, and evaluate the activation and functionality of CART cells through ELISA or Cytokine Bead Array.
A: CART therapy evaluation: For evaluating the effectiveness of novel anti-CD20 CART therapies, including comparison and optimization of different CAR constructs. Drug screening: Screen and validate drugs that enhance CART cell function, and evaluate the effects of these drugs on CART cell expansion, persistence, and anti-tumor activity. Safety testing: Test the safety of CART cell therapy in the pre-clinical stage, evaluate potential off-target effects and cytokine storm risks, and ensure the safety and effectiveness of the treatment.
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The cell quality is very high, showing excellent stability and consistent growth during culture. In particular, the expression of anti-CD20 CAR is very stable and can accurately identify and target CD20-expressing cells, which is very helpful for my experiments. This cell line has played an important role in immune research and is one of the best cells I have ever used.
The customer service staff were professional and enthusiastic. They answered all my questions in detail and helped me choose the most suitable product. After the cell line arrived, its performance in culture was exactly as described, especially the stable expression of the anti-CD20 CAR and its efficient targeting ability, which was exactly as advertised. The cells also perform very well in suspension and adherent culture, which enhances my sense of trust. As a researcher, I value product reliability very much and I was very satisfied with this purchase experience.
During use, the cell growth parameters such as proliferation curves and gene expression levels were very consistent, which greatly improved my work efficiency.
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