November 2021 Share
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.
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 10 minutes.
Most immune cells are isolated from whole blood samples in living organisms. Two common host subjects for sample extraction are humans and mice. 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. This process must be performed in both species. While the protocols for human RBC depletion and mouse RBC depletion are virtually the same, they have different implications for what they allow.
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. In humans, red blood cells make up roughly 40-45 percent of blood. When working in a blood-derived starting material, residual RBC contamination is a common concern as it can be challenging to remove all of these cells.
Human red blood cells can be removed from blood-derived samples in a multitude of ways. Samples are frequently centrifuged to sort cells by density or size, which can be one method to remove the bulk of the RBCs from the sample, but it is not uncommon for residual RBC contamination to remain.
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.
Mouse immune cell isolation protocols often use murine splenocyte samples. The mouse spleen has a higher concentration of T lymphocytes than most other organs in the body; this makes it an ideal subject for immune cell separation assays. To free up the lymphocytes for isolation, the spleen tissue is dissociated to release the cells from the tissue. Mouse RBC depletion allows researchers to clear the contaminating red blood cells from the splenocyte sample.
Along with humans, mice also have an adaptive immune system that allows them to respond to and immunize against harmful pathogens. While there are slight differences in the complexities between mouse and human immune processes, the two species share many cell types with one another. This means that murine cells can often be used as a reference for human cells. The accessibility and ease of mouse cell isolation make it a common research alternative to human cell isolation.
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