Cell & Gene Therapy
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Anne Lodge, PhD
What Are Irradiated PBMCs?
This blog post has been revised April 21, 2023 to reflect updates in scientific data.
Sometimes researchers need access to the components and benefits of normal peripheral blood mononuclear cells, but don’t want the cells to proliferate in their experiments. That’s where irradiated PBMCs come in handy.
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What Makes PBMCs “Irradiated”?
Irradiated PBMCs are normal PBMCs that have been exposed to radiation from an X-ray source to create enough DNA damage that the cells can no longer divide. The irradiated cells are still alive and can produce cytokines and other soluble factors, but are unable to proliferate.
How Should Irradiated PBMCs Be Used?
Irradiated PBMCs are ideal for use as antigen-presenting cells in combination with anti-CD3 antibodies to stimulate T cell activation and proliferation. Since they cannot proliferate, there is no risk of the PBMCs contaminating the cells of interest.
Irradiated PBMCs may be used as a feeder layer to provide metabolites to support the growth of the cells of interest.
What Are the Drawbacks of Irradiated PBMCs?
One drawback of irradiated PBMCs is that they are single-use reagents. They definitely will not grow in a culture.
What is an Alternative to Irradiated PBMCs?
If you do not wish to use PBMCs exposed to radiation, PBMCs can be treated with mitomycin C instead to achieve a similar effect. Mitomycin C is a naturally occurring DNA crosslinking agent and also prevents cell division.
The downside of mitomycin C is that it involves incubation and multiple washes to ensure there is no residual mitomycin C carried over to the cells in your culture. During the incubation and washing process, as much as half of the starting PBMCs can be lost.
In addition to the extra handling, some labs prefer not to use the toxic chemical mitomycin C and instead purchase ready-to-thaw-and-use irradiated PBMCs.