How Do You Culture and Expand T Cells, PBMC, and Other Immune Cells?
We get these types of culture questions all the time. After all, we developed the immune cells, have tested their functionality, and created research protocols specific to them. So naturally, we can help!
However, the answers to these questions aren’t always straightforward. Our responses often depend on each researcher’s specific lab conditions, product handling, research criteria, and experience level.
Below we’ve compiled some of our most popular questions related to cell culture and expansion along with our responses. These should get you headed in the right direction.
My PBMC culture turned into all T cells. What happened to the other cell types?
Human PBMC are a mixture of different cell types, including T cells, B cells, monocytes, and natural killer (NK) cells, none of which will grow without stimulation (and monocytes will never proliferate).
Depending on the stimulus you use, your PBMC culture will become dominated by T cells or B cells, quickly turning the cell mixture into a single cell type. With that understanding, feel free to grow your cell type of interest, just don’t expect the expanded cells to behave like PBMC after culture.
How many cells can we expect to generate from a culture of T cells?
Your expected results depend on several factors, including culture conditions and stimulus. If you stimulate your culture with CD3 and anti-CD28, you may get 1.5X as many T cells as you had in your original PBMC. Adding cytokines like IL-2 can dramatically increase that number.
Can we culture and amplify your antigen-specific T cells?
Growing T cells is our specialty, especially as we are the only commercial source of antigen-specific T cells. Culturing and expanding antigen-specific T cells is a complex process, so we recommend trusting your cultures to our immunology CRO team. That way, you’ll know that the T cells you receive from us will be active in culture.
My monocytes are not expanding. What did I do wrong?
Monocytes can differentiate into dendritic cells, macrophages, or osteoclasts (get the protocol here), but they do not proliferate. Don’t expect to generate more cells during culture.
Do you culture human monocytes in any particular type of flask? I find macrophages very difficult to remove from TC-treated and non-TC-treated flasks.
Macrophages are indeed difficult to move. If we intend to move them, we use Corning Ultra-Low Attachment cultureware. Otherwise, we culture them where we want them.
What is the best cell culture medium?
There are many factors to consider when selecting a culture medium. For an in-depth exploration of the effects of culture medium on experimental outcomes, read our case study.