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Pay attention to these three points to make your 4T1 cells more “beautiful”

Source: PricellaPublished: 2024-04-22

4T1 is a 6-thioguanine resistant cell line selected from the 410.4 tumor without mutagen treatment. 4T1-induced tumors can serves as both post-operative and non-surgical models because they metastasize spontaneously in both models with similar kinetics. This article summarizes the characteristics of 4T1 cells and the precautions of reception and cultivation, facilitating a smooth start and successful experiments for researchers.

Figure 1. Normal growth morphology of 4T1 cells

Cellular characteristics

4T1 breast cancer is a tumor with low immunogenicity and high malignancy. After the primary breast tumor grows, it can quickly and spontaneously metastasize to lymph nodes, lungs, liver, brain and blood [1].

4T1 cells derived from BALB/c mice are very close to human breast cancer in terms of immunogenicity, metastasis and growth characteristics [2].

4T1 growth characteristics are similar to highly invasive human metastatic breast cancer. During surgery, the disease degree is equivalent to that of human stage IV breast cancer, and 4T1 breast cancer in mice with poor immunogenicity can be established as a postoperative animal model[3].

Precautions of receiving the cells

Place the cells in the incubator for 2~4 hours to observe their status.

If there is a significant amount of cell shedding, the original bottle culture medium can be centrifuged and collected for precipitation. After digestion and treatment, it can be combined with the adherent cells of the original bottle cells and inoculated.

If there is no shedding of cells basically, about 80% density can be passaged at a ratio of 1:3 to 1:4, and 100% density can be passaged at a ratio of 1:5.

Precautions of culturing the cells

1. The growth rate of this cell is fast, and the higher the cell density, the more difficult it is to digest. It is recommended to conduct digetion in stages. Once digested until there is cell movement and floating, the remaining cells should be digested again to form single cells as much as possible. It is normal for a small amount of cell clusters to adhere to the wall and not disperse on the second day of cell passage. With continued culture, these clusters will disperse.

2. If there is a phenomenon of cell rounding during cultivation, it may be due to excessive density, which prevents the cells from despersion. This can be improved by reducing the passage density.

3. Recommended growth medium for 4T1 cells: 4T1 Cells Complete Medium (Cat. No.: CM-0007)


[1] Pulaski BA, Clements VK, Pipeling MR, Ostrand-Rosenberg S. Immunotherapy with vaccines combining MHC class II/CD80+ tumor cells with interleukin-12 reduces established metastatic disease and stimulates immune effectors and monokine induced by interferon gamma. Cancer Immunol Immunother. 2000 Apr;49(1):34-45.

[2] Pulaski BA, Ostrand-Rosenberg S. Reduction of established spontaneous mammary carcinoma metastases following immunotherapy with major histocompatibility complex class II and B7.1 cell-based tumor vaccines. Cancer Res. 1998 Apr 1;58(7):1486-93.

[3] Pulaski BA, Terman DS, Khan S, Muller E, Ostrand-Rosenberg S. Cooperativity of Staphylococcal aureus enterotoxin B superantigen, major histocompatibility complex class II, and CD80 for immunotherapy of advanced spontaneous metastases in a clinically relevant postoperative mouse breast cancer model. Cancer Res. 2000 May 15;60(10):2710-5.

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