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How Does RBC Depletion Work?

November 2021

Sample preparation is often a critical first step in any number of life science workflows. Due to the inherent complexity within samples and diversity between applications, sample preparation needs can vary widely. One common first step is a “cleanup” step to deplete residual red blood cells (RBCs) from the sample before proceeding with downstream analysis. Once the RBCs have been depleted, the resulting blood product can be used across many cell research and immunotherapy applications. 

Red blood cell depletion is a method of selection that removes the red blood cells from a sample. Depending on the specific technique used, red blood cells can be removed with very high precision in very short amounts of time. For example, Akadeum’s Human Red Blood Cell Depletion Microbubbles can remove 99% of RBC contamination in only 20 minutes with no significant effects on the white blood cells. However, other methods can have damaging off-target effects. Protocols for lysing of RBCs, for example,  can inadvertently harm other cell types in the sample in ways that effect downstream results.

Human RBC Depletion 

Immune cells are typically isolated from whole blood samples in living organisms. When attempting to study lymphocytes or any other specialized cell that is not a red blood cell, RBCs frequently need to be removed from the sample as they create background “noise” that can get in the way of further analysis or slow down further cell sorting methods such as fluorescence-activated cell sorting (FACS). 

Human red blood cell depletion serves a variety of purposes. With red blood cells being the most abundant subset of cells in blood samples, they can easily get in the way when trying to isolate other cell populations. Red blood cells make up roughly 40-45 percent of the blood by volume in humans. When working with a blood-derived starting material, residual RBC contamination is a common concern as it can be challenging to remove all of these cells and not damage the overall health and function of the other cells in the sample.

Human red blood cells can be removed from blood-derived samples in many ways. One method for removing the majority of RBCs from blood samples involves centrifugation to sort cells by density or size. Unfortunately, it is not uncommon for residual RBC contamination to remain and white blood cells to become damaged. This can have a significant effect on the data generated using these cells in downstream applications.

Human RBC depletion allows scientists to reduce red blood cell contamination in a sample, enabling accurate and efficient cell sorting. Using microbubbles from Akadeum for RBC cleanup before beginning the cell sorting process can not only reduce sort times by 1/3 or more but can result in a larger population of healthy, viable cells at the end of the sort. This is much more difficult to obtain using other methods, such as density centrifugation or lysis of RBCs. 

Increase RBC Depletion Efficiency with Akadeum

RBC Depletion is a crucial preparation step in isolating viable blood cells for research. With Akadeums Microbubble Depletion technology, cell separation has never been easier. Once the microbubbles are mixed into the blood sample, they will bind to the contaminating RBCs. Using the innate buoyant nature of the microbubbles, the RBCs float to the top of the sample to be discarded. This protocol takes place in less than twenty minutes and can remove up to 99% of the RBC contaminants. Learn more about Akadeum’s Human Red Blood Cell Depletion Microbubbles.

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