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Cat.No. : CSC-RR0224 Host Cell: 4T1
| Cat. No. | CSC-RR0224 |
| Description | Luc Reporter Cell Line-4T1 is engineered to stably overexpress luciferase reporter gene. It is an ideal positive control for use in bioluminescence assays. In addition, 4T1 cells form tumors post implantation into immunocompromised or syngenic mice. The in vivo growth of these tumors can be monitored using bioluminescent imaging. |
| Gene | Luciferase |
| Host Cell | 4T1 |
| Host Cell Species | Mus musculus (Mouse) |
| Stability | Validated for at least 10 passages |
| Application | 1. Gene expression studies 2. Protein localization 3. Drug screening and toxicology 4. Live cell imaging |
| 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 |
Hypoxia is one of cancer's hallmarks. Using a miR-210-specific reporter system, the researchers observed the transfer of miR-210 from hypoxic breast cancer cells to surrounding cells. Exosomes containing miR-210 were transported throughout the tumor microenvironment, according to both in vitro and in vivo studies. MiR-210 modulated the expression of vascular remodeling genes such as Ephrin A3 and PTP1B, encouraging angiogenesis. This study demonstrates how exosome-mediated communication promotes the transmission of miRNAs from hypoxic cancer cells to neighboring cells, which influences tumor growth.
Figure 1. The researchers designed a miR-210 reporter construct to image miR-210 expression in 4T1-luc cells. Luciferase signals from these cells were modulated by miR-210 binding. Bioluminescence imaging and luciferase assays demonstrated dose-dependent signal reduction upon DFO treatment (400 μM). In vivo imaging of 4T1/miR-210 tumors confirmed decreased luciferase signals post-DFO injection, correlating with altered HIF-1α expression. These findings highlight the utility of 4T1-luc2/miR-210 cells in studying miR-210 regulation under hypoxic conditions. (Jung KO, et al., 2017)
A: The Luc Reporter Cell Line-4T1 is optimally utilized in cancer research, specifically for studying breast cancer metastasis and tumor immunology. It offers the significant advantage of allowing real-time, non-invasive monitoring of tumor progression and metastasis in live animals through bioluminescence imaging. This capability enables researchers to quantitatively measure tumor burden over time without euthanizing the animals, facilitating longitudinal studies on tumor growth and regression in response to treatments.
A: Key considerations include ensuring consistent luciferin substrate administration, maintaining the animals under appropriate conditions to avoid stress-induced variability, and standardizing imaging protocols (e.g., timing post-luciferin injection and camera settings). Additionally, it's vital to use an appropriate control group and calibrate the imaging system to account for background luminescence and signal attenuation due to tissue depth.
A: By incorporating the Luc Reporter Cell Line-4T1 into animal models, researchers can non-invasively track tumor response to therapeutic agents in real-time. This approach allows for the dynamic assessment of drug efficacy, enabling the identification of the most promising therapeutic agents for further development. The implications for personalized medicine are profound, as this methodology can be adapted to screen for drugs that are most effective against the specific genetic makeup of a patient's tumor, potentially leading to more targeted and effective treatments.
A: Potential limitations include the challenge of detecting very small metastatic sites due to the sensitivity limits of bioluminescence imaging, and the potential for variability in luciferase expression among cells. To mitigate these issues, researchers can employ imaging systems with enhanced sensitivity, use multimodal imaging approaches to validate findings, and perform rigorous quantification of luciferase expression levels to ensure consistency. Additionally, advancing the imaging technology and developing more sensitive luciferase substrates could further improve detection capabilities.
A: When designing experiments to explore the tumor microenvironment with the Luc Reporter Cell Line-4T1, it's crucial to consider co-culturing the luciferase-expressing 4T1 cells with other cell types relevant to the tumor microenvironment, such as immune cells or fibroblasts. This can help elucidate interactions between tumor cells and their surroundings. Additionally, employing imaging techniques that can spatially resolve the bioluminescent signal within the tumor can provide insights into how different regions of the tumor microenvironment influence cancer progression. Combining the Luc Reporter Cell Line-4T1 with other reporter systems that can mark different cell types or physiological states (e.g., hypoxia) can also enrich the data and provide a more comprehensive understanding of the tumor microenvironment.
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The 4T1 Luc Reporter Cell Line provides a highly sensitive and quantitative measure of reporter gene expression, which is valuable for assessing subtle changes in gene regulation.
United States
11/24/2023
The stable nature of the luciferase reporter in 4T1 cells ensures a uniform response, which is critical for obtaining consistent and reliable data across experiments.
Canada
07/22/2021
The 4T1 Luc Reporter Cell Line has minimal background signal, which reduces noise and increases the dynamic range for detecting luciferase activity.
French
07/19/2022
The 4T1 Luc Reporter Cell Line can be used to assess transfection efficiency and optimize transfection protocols, which is essential for gene function studies.
United States
09/10/2022
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