Cat.No. : CSC-RR0539 Host Cell: MC38
| Cat. No. | CSC-RR0539 |
| Description | This cell line is engineered to stably overexpress GFP and luciferase reporter genes. MC38 is a a mouse colon adenocarcinoma cell line. This cell line is a useful tool for both fluorescent and bioluminescent tracking of MC38 cells. |
| Gene | GFP/Luc |
| Host Cell | MC38 |
| Host Cell Species | Mus musculus (Mouse) |
| Stability | Validated for at least 10 passages |
| Reporter Type | Fluorescent protein |
| 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 |
The researchers investigated how maternally expressed gene 3 (MEG3) combats chemoresistance in acute myeloid leukemia (AML). They discovered that MEG3 and microRNA (miR)-493-5p are downregulated in resistant AML cells when compared to parental cells. MEG3 increases miR-493-5p levels, which target methyltransferase-like 3 (METTL3). METTL3, in turn, stimulates MYC expression by increasing its m6A alteration. MEG3 and miR-493-5p overexpression, as well as METTL3 deletion, decreased cell growth and increased apoptosis in HL-60 and Molm13 cell lines, respectively. MEG3 also boosted AML cells' sensitivity to arabinocytosine (AraC). Suppressing miR-493-5p reversed the effects of MEG3. These data indicate that MEG3 increases AML cell chemosensitivity by modulating the miR-493-5p/METTL3/MYC axis.
Figure 1. The researchers confirmed that METTL3 is a miR-493-5p target by doing dual luciferase reporter tests with the MOLM13 cell line. They measured METTL3 expression in distinct AML and normal samples and discovered substantial differences. (Wang A, et al., 2022)
A: The GFP/Luc Reporter Cell Line - MC38 utilizes a dual reporter system where the green fluorescent protein (GFP) serves as a visual marker and firefly luciferase (Luc) as a quantitative reporter. This system enables both visualization of GFP-expressing cells under fluorescent microscopy and quantification of luciferase activity through bioluminescence assays.
A: In studying tumor progression in vivo, the GFP/Luc Reporter Cell Line - MC38 can be injected into animal models. The GFP fluorescence allows tracking of tumor cells over time using imaging techniques, while luciferase activity provides a quantitative measure of tumor growth, metastasis, and response to treatments via bioluminescence imaging.
A: The GFP/Luc Reporter Cell Line - MC38 offers several advantages over traditional cell lines. Firstly, the dual reporter system allows for both qualitative visualization and quantitative analysis of tumor cells in vivo and in vitro. Secondly, the stable expression of GFP and luciferase ensures consistent and reliable results over time. Additionally, the MC38 cell line, derived from murine colon carcinoma, mimics aspects of human colorectal cancer, making it relevant for translational research.
A: By utilizing the GFP/Luc Reporter Cell Line - MC38 in co-culture experiments with immune cells or stromal cells, researchers can investigate the dynamic interactions within the tumor microenvironment. The GFP fluorescence allows tracking of tumor cells, while luciferase activity enables quantification of changes in tumor growth or response to immunotherapies or other interventions.
A: Generating stable cell lines with the GFP/Luc Reporter Cell Line - MC38 involves transfecting the cells with plasmids containing the GFP and luciferase genes, followed by selection with appropriate antibiotics or selection markers. Single-cell cloning can be performed to isolate individual clones with stable expression of both reporters. These clones can then be expanded and characterized for their GFP fluorescence and luciferase activity before experimental use.
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The GFP/Luc Reporter Cell Line - MC38 includes both GFP (Green Fluorescent Protein) and luciferase as reporters. This dual-reporter setup allows for versatile experimental design, enabling both real-time fluorescence imaging and bioluminescence assays. We can observe cellular events in live cells using GFP and quantify promoter activity with luciferase in the same cell population, enhancing experimental flexibility.
Equipped with GFP, the GFP/Luc Reporter Cell Line - MC38 allows for continuous live-cell imaging. This capability is crucial for monitoring cellular processes in real time, such as migration, proliferation, and drug responses, without the need to terminate the assay to assess outcomes, preserving the integrity of the experimental timeline.
The luciferase component of the GFP/Luc Reporter Cell Line - MC38 makes it suitable for high-throughput screening (HTS) applications. Luciferase assays can be rapidly conducted with automated systems, allowing for the efficient screening of large compound libraries or genetic screens, which is beneficial for drug discovery and gene function studies.
This cell line enables the collection of both quantitative (from luciferase activity) and qualitative (from GFP fluorescence) data. We can thus obtain robust datasets that offer insights into both the magnitude and the visual context of cellular responses, providing a comprehensive view of experimental variables.
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