Mouse Cd47 Knockout Cell Line-MC38
Cat.No. : CSC-RT2698
Host Cell: MC38 Target Gene: Cd47
Size: 1x10^6 cells/vial, 1mL Validation: Sequencing
Inquire for Price
Cell Line Information
Cell Culture Information
Safety and Packaging
| Cat. No. | CSC-RT2698 |
| Cell Line Information | This cell is a stable cell line with a homozygous knockout of mouse Cd47 using CRISPR/Cas9. |
| Target Gene | Cd47 |
| Host Cell | MC38 |
| Size Form | 1 vial (10^6 cell/vial) |
| Shipping | Dry ice package |
| Storage | Liquid nirtogen |
| Species | Mouse |
| 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 |
Inquiry
Background
Case Study
Applications
CD47 is a transmembrane protein that is widely expressed on various cells, including those in the immune system, making it a key player in cell interactions and immune responses. CD47, also known as integrin-associated protein (IAP), belongs to the immunoglobulin superfamily and is involved in different physiological and pathological processes. The protein interacts with signal regulatory protein α (SIRPα) on the surface of macrophages, inhibiting their ability to engulf and destroy cells. This interaction is essential for maintaining self-tolerance, preventing the immune system from attacking its own cells. Cancer cells are known to exploit this mechanism by overexpressing CD47, thereby escaping immune surveillance and destruction. This has made CD47 an interesting target for cancer research, leading to the development of therapeutic agents designed to block the CD47-SIRPα interaction, thereby enhancing the ability of the immune system to target and eliminate tumor cells.
In addition to its anti-phagocytic role, CD47 is involved in regulating other cellular processes such as apoptosis, proliferation, migration, and adhesion. Studies have shown that it is involved in regulating integrin signaling, which is essential for cell motility and stability. This makes CD47 an integral part of various physiological processes, including development, homeostasis, and wound healing. Recent studies have also implicated CD47 in diseases other than cancer. For example, its dysregulation has been observed in autoimmune diseases, cardiovascular diseases, and neurodegenerative diseases.
CD47 on tumor cells prevents phagocytosis, whereas PD-L1 inhibits T cell-mediated tumor killing. However, whether and how CD47 and PD-L1 coordinate is unclear. Studies here found that CD47 and PD-L1 on tumor cells synergistically inhibit innate and adaptive sensing to evade immune control. Targeted blockade of CD47 and PD-L1 on tumor cells using bispecific anti-PD-L1-SIRPα showed significantly enhanced tumor targeting and therapeutic efficacy over monotherapy. Mechanistically, systemic delivery of the dual-targeting heterodimer significantly increased DNA sensing, DC cross-presentation, and antitumor T cell responses. In addition, chemotherapy that increased the "eat me" signal further synergized with the bispecific agent to better control the tumor. These data suggest that tumor cells evolve to exploit innate and adaptive checkpoints to evade antitumor immune responses and that tumor cell-specific dual targeting of both checkpoints represents an improved strategy for tumor immunotherapy.
To test whether CD47 is required for tumor cell immune evasion, the researchers generated a CD47-deficient colorectal tumor cell line, named "MC38-CD47 knockout" (MC38-CD47KO). MC38-CD47 knockout tumor cells grew as well as parental cells in vitro. When inoculated subcutaneously into immune-competent wild-type (WT) animals, their growth was slower than that of parental cells, but not completely stopped (Figure 1A). This result suggests that CD47 plays only a partial role in tumor cell immune evasion and that other mechanisms act together or independently of CD47 in tumor cell immune evasion. Previous studies have shown that MC38 growth is controlled by adaptive T cells and that T cell depletion enhances tumorigenesis. Similarly, MC38-CD47KO growth was also controlled by T cells (Figure 1B). Consistent with this finding, PD-L1 expression was upregulated on MC38-CD47KO tumor cells under T cell pressure (Figure 1C). The researchers further knocked out PD-L1 on MC38-CD47 knockout cells to obtain MC38-CD47/PD-L1 double knockout cells (MC38-CD47/PD-L1DKO). MC38-PD-L1 knockout cells were also generated as a control. In sharp contrast to MC38-CD47KO and MC38-PD-L1KO cells, MC38-CD47/PD-L1DKO cells almost completely stopped growing (Figure 1D). The growth arrest of MC38-CD47/PD-L1DKO was dependent on host CD8+ T cells, as their growth levels in Rag1−/− mice or WT mice lacking CD8+ T cells were comparable to WT MC38 tumor cells (Figures 1E and 1F). Together, these data suggest that CD47 and PD-L1 on tumor cells act in concert to evade tumor immune escape.
Figure 1. CD47 and PD-L1 on Tumor Cells Coordinate to Evade Immune Control. (Liu X, et al., 2018)
The mouse CD47 knockout cell line - MC38 is a valuable tool for biomedical research, primarily used to explore the role of CD47 in cancer biology and immunotherapy. CD47 is often referred to as a "don't eat me" signal that acts as an inhibitory signal for macrophage-mediated phagocytosis. Here are some key applications of this knockout cell line:
Cancer Immunotherapy Research: The MC38 cell line with the Cd47 gene knocked out is widely used in cancer immunotherapy research. By knocking out Cd47, researchers can study how the absence of this signal affects tumor growth and response to immune checkpoint inhibitors. This can reveal new therapeutic targets and enhance the efficacy of existing cancer treatments.
Drug Development: In drug research, MC38 Cd47 knockout cells can be used to screen for novel drugs that target the CD47-SIRPα interaction pathway. By understanding how these drugs affect cancer cells that lack Cd47, researchers can develop more effective therapies that enhance the immune system's ability to destroy cancer cells. This also helps evaluate the safety and efficacy of potential new drugs.
Tumor Immune Evasion Mechanisms: Studying Cd47 knockout MC38 cells allows researchers to delve into the mechanisms by which tumors evade the immune system. Because CD47 plays a critical role in protecting cells from immune system attack, its absence can provide insight into alternative immune evasion pathways and help identify additional potential therapeutic targets within the tumor microenvironment.
Biomarker Studies: The MC38 Cd47 knockout cell line is a valuable tool for biomarker discovery and validation. By comparing differences in immune responses and tumor progression between wild-type and Cd47 knockout cells, researchers can identify biomarkers that predict response to immunotherapy.
For research use only. Not intended for any clinical use.
