4T1-OVA Luciferase Reporter Cell Line

Frequently Asked Questions

• What is the 4T1-OVA Luciferase Reporter Cell Line?

  The 4T1-OVA Luciferase Reporter Cell Line is derived from the 4T1 cell line, which is a mouse breast cancer cell line. This reporter cell line has been engineered to express both the ovalbumin (OVA) gene and the luciferase gene, allowing for the tracking of tumor growth and immune response dynamics through bioluminescent imaging. This makes it valuable for cancer research, particularly for evaluating cancer immunotherapies in mouse models.

• What is the recommended culture medium for the 4T1-OVA Luciferase Reporter Cell Line?

  The 4T1-OVA Luciferase Reporter Cell Line should be cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS). To maintain selective pressure and ensure stable expression of the luciferase and OVA genes, the culture medium should also contain 4.0 µg/ml of puromycin.

• What are the applications of the 4T1-OVA Luciferase Reporter Cell Line?

  This cell line is primarily used for studying tumor immunology, oncology, and the efficacy of immunotherapeutic agents. Its ability to express luciferase enables real-time non-invasive imaging of tumor progression and regression in vivo, facilitating studies on how immune cells interact with tumors and respond to immunotherapies.

• How was the 4T1-OVA Luciferase Reporter Cell Line generated?

  The 4T1-OVA Luciferase Reporter Cell Line was generated by transducing the original 4T1 cells with lentiviral particles containing a construct for the expression of luciferase and the chicken ovalbumin gene. This was followed by selection with puromycin to ensure the establishment of a cell population stably expressing both genes.

• What validation tests confirm the expression of the genes?

  The expression of the ovalbumin gene in the 4T1-OVA Luciferase Reporter Cell Line was confirmed through RT-qPCR analysis. This testing demonstrated efficient expression of the ovalbumin mRNA in the cell line compared to wild-type 4T1 cells, where this gene was undetectable. This validation confirms the successful integration and functional expression of the transgene in the engineered cell line.