Cat.No. : CSC-RO0610 Host Cell: CT26
| Cat. No. | CSC-RO0610 |
| Description | This cell line is derived from CT26 and is engineered to stably overexpress Human CD20. |
| Gene | CD20 |
| Gene Species | Homo sapiens (Human) |
| Host Cell | CT26 |
| Host Cell Species | Mus musculus (Mouse) |
| 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 |
A: When utilizing the Human CD20 Stable Cell Line - CT26 to evaluate anti-CD20 monoclonal antibodies' efficacy, it's essential to include both negative and positive controls to validate the assay's specificity and sensitivity. A non-transfected CT26 cell line should serve as a negative control, whereas a well-characterized anti-CD20 antibody can be used as a positive control. Optimizing antibody concentrations and incubation times is critical for distinguishing between effective and ineffective antibodies. Flow cytometry should be used to quantitatively assess CD20 expression and antibody binding, ensuring cells are properly stained and analyzed under consistent conditions.
A: To investigate the mechanisms of action of CD20-targeted therapies using the Human CD20 Stable Cell Line - CT26, researchers can perform a series of assays that dissect the pathways involved in drug action. This includes assessing cell viability post-treatment using assays such as MTT or Annexin V/PI staining to determine apoptotic and necrotic cell death. Co-culturing the cell line with immune effector cells, such as natural killer cells, can elucidate antibody-dependent cellular cytotoxicity (ADCC) mechanisms. Additionally, complement-dependent cytotoxicity (CDC) assays can be performed by adding human complement serum post-antibody treatment. These experiments provide insights into how CD20-targeted therapies induce tumor cell death.
A: To verify the stable expression of CD20 in the Human CD20 Stable Cell Line - CT26 over extended culture periods, periodic validation using flow cytometry is recommended. This involves staining the cells with a fluorescently labeled anti-CD20 antibody and analyzing the percentage of CD20-positive cells. Quantitative PCR (qPCR) can also be used to measure the transcript levels of CD20, ensuring the gene's expression remains consistent. It's advisable to perform these checks at various passages to monitor for any potential drift in CD20 expression. Additionally, Western blotting can be utilized to assess the protein levels of CD20, providing a comprehensive view of its expression stability.
A: When adapting the Human CD20 Stable Cell Line - CT26 for in vivo studies in murine models, several considerations must be addressed. First, ensure the murine model is immunocompromised to prevent immune rejection of the humanized cells. Selecting an appropriate model, such as NSG or NOD-SCID mice, is critical for successful engraftment. Secondly, monitoring for potential immunogenicity of the human CD20 expressed on CT26 cells is essential, as it might elicit an immune response. Additionally, establishing optimal cell inoculation numbers and routes of administration (e.g., subcutaneous, intravenous) is crucial for replicating disease pathology accurately. Lastly, ethical considerations and institutional guidelines for animal care and use must be strictly followed.
A: Assessing the impact of genetic background on the expression and function of CD20 in the Human CD20 Stable Cell Line - CT26 requires a comparison with other cell lines or the same cell line with different genetic backgrounds. This can be achieved by introducing specific genetic modifications using CRISPR/Cas9 or other gene-editing tools to alter genes that may interact with CD20 expression or function. Subsequent analyses, including flow cytometry for CD20 surface expression and functional assays such as ADCC or CDC, can elucidate the role of these genetic factors. Additionally, RNA sequencing (RNA-Seq) could be employed to identify differential gene expression profiles that may influence CD20's role in these cell lines.
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The Human CD20 Stable Cell Line - CT26 maintains consistent expression of the CD20 protein, crucial for reliable experimental outcomes. This consistent expression ensures that the results of studies conducted using this cell line, particularly those involving the behavior and interaction of CD20, are reproducible and dependable.
Stability is a key feature of the Human CD20 Stable Cell Line - CT26, where the cell line shows high resilience and uniformity over extended cultures. This stability is important for conducting experiments that require long-term cell growth without the loss of phenotype or function.
This cell line is optimized for testing and developing anti-CD20 antibodies, a common therapeutic approach in conditions like lymphoma and leukemia. The Human CD20 Stable Cell Line - CT26 provides a robust platform for evaluating the efficacy of antibodies in binding and neutralizing CD20, crucial for therapeutic antibody development.
The Human CD20 Stable Cell Line - CT26 exhibits high transduction efficiency, making it easier to introduce additional genetic elements if needed. This feature is particularly useful for us looking to study gene function in the context of CD20 expression or to create more complex disease models.
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