Antigen-presenting dendritic cells are an important factor in studying the immune system. If you already have dendritic cells (DC) in your lab, you can culture them to achieve a desired phenotype and activate them to recognize immune or inflammatory responses.

 

5 Tips for Dendritic Cell Culture and Activation

1. Avoid Transferring DC During Culture

Dendritic cells are composed of both adherent and non-adherent cells, making the movement of cells from one culture vessel to another a dicey situation. Those adherent cells won’t come off the plastic without a real fight. When possible, set up your cells in the culture configuration you want to use in your experiment, so you don’t have to disturb the cells.
If you do switch vessels during your culture or experimentation, expect to recover fewer cells.

2. Add the Right Proportion of Cytokines to Preserve the DC Phenotype

Dendritic cells will survive in culture for approximately one week. If you have plans to maintain your dendritic cells after an overnight culture, you need to add the proper proportion of granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin 4 (IL4) recombinant human proteins.

We recommend adding 500 U/mL of GM-CSF and 500 U/mL of IL4, which you can purchase from suppliers like Thermo Fisher or PeproTech. This will ensure your dendritic cells maintain their phenotype after thawing.

3. Activate Your DC, If Necessary

Our dendritic cells are immature, but you can easily activate or mature them by adding lipopolysaccharides (LPS), poly(I:C), or other pathogen pattern molecules. Alternatively, you could use cytokines, such as IFNγ or TNFα, in combination with prostaglandin to achieve a similar result.

Depending on the stimuli used, you can get a slightly different dendritic cell that could suppress an immune response rather than ramp it up.

4. Always Include Serum When Stimulating DC with LPS

You can stimulate your dendritic cells with LPS, but you must include serum as well for proper activation. The soluble CD14 in the serum facilitates binding of the LPS to the dendritic cells.

For serum, you can use human serum or fetal bovine serum to achieve the desired result.

5. Follow Our Detailed Protocol for Successful DC Cultures

In addition to these tips, be sure to follow the procedures outlined in our Human Dendritic Cell Culture protocol.
Have other research questions? Ask a scientist and get a quick answer!
 

2 Comments

  1. Hi, thank you for these tips. I want to ask something. As far a I know we couldn’t make cryopreserved storage of dendritic cells. Than how long can we culture these cells by changing the media?

    • If you are starting from monocytes, it takes 4-6 days before they fully differentiate into DC (when culturing with GM-CSF and IL-4). This is the best time to harvest the DCs for your experiment. You can add fresh media to maintain them in culture for 2-3 more days. However, these cells will be increasingly adherent to tissue culture plastic.

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