Individual cell study provides insight into their specific functions and roles within the human body. Knowing exactly what certain cells do enables scientists to harness and learn from them. Cell separation is a major driving force behind the push for personalized medicines and the ability to treat large populations with effective generalized methods.
What are the Approaches to Isolating Cells?
Cell separation methods typically take one of the three approaches listed below:
- Positive selection
- Negative selection
- Depletion
The method you use should be determined by the context of your experiment.
Positive Selection Cell Separation
When the cell type of interest is targeted by the removal mechanism and retained for downstream applications, this is referred to as positive selection. This method involves targeting the desired cell population with an affinity molecule specific to the cell’s surface marker, leaving unwanted cells in the sample.
Cell Separation by Negative Selection
Negative selection occurs when undesirable cell types are labeled with affinity molecules such as antibodies or proteins that target specific cell markers or populations and then removed, leaving only one cell type unaffected. The untouched cell sample is then collected for later use.
Positive vs Negative Selection
The choice between positive and negative selection will be heavily influenced by the context of the experiment. Positive selection can provide higher purity than negative selection if the target cell has a very clear selection marker on its surface. If your selection markers are unclear and you plan to perform downstream assays on your isolated cells, negative selection will remove unwanted cells faster without affecting the enriched population.
Cell Depletion
Cell depletion is the third and simplest method for removing a single cell type from a biological sample. This method is commonly used to remove large amounts of a single common contaminant, such as red blood cells (RBCs) or dead cells. RBC depletion kits can be used to further purify a sample if it is heavily saturated with residual RBCs after the cell separation process.
Cell Separation Applications
There are numerous cell separation methods, but the potential uses of isolated cells vastly outnumber the methods. A variety of scientific disciplines can benefit from healthy, purified cell samples. Some of the applications that can be performed or carried out with cell separation or isolated cells are listed below.
Blood Separation – PBMC Isolation from Buffy Coat
When working with whole blood samples, white blood cells (WBCs) and platelets make up a very small percentage of the cells. After centrifuging blood and determining how to separate plasma from blood, these cells form a thin layer known as a buffy coat, which can be separated for research purposes. The buffy coat’s high concentration of peripheral blood mononuclear cells (PBMCs) makes it ideal for studying how the body responds to infectious diseases and dangerous pathogens.
Read More: What Is PBMC? Human PBMC Cells, PBMC Composition, and Isolation Tools
PBMCs are immune cells that have been isolated for research or medical treatment. A person’s immune system can be boosted by receiving PBMCs via transfusion.
RBC Depletion in a Cell Sample
The most common contaminant in PBMC isolation from whole blood is residual RBCs. These cells do not function the same as WBCs and, if present in the sample, can impede research into how immune cells behave. Cell separation can be used to easily remove these cells and clean a sample for subsequent analysis and applications.
T Cell Separation for Research and Practical Application
Using cell separation techniques to isolate T cells opens up a world of possibilities for immunology research and treatment. Understanding the different types of immune cells in the human body can help guide medical research and provide insight into the immune response.
CAR T Cell Isolation for CAR T Cell Therapy
The receptors found on the surface of cancerous cells are known as chimeric antigen receptors (CARs). Not all T cells can recognize these specific antigens. Scientists can use cell separation to accomplish two things:
- Isolate cancer-targeting cells, culture them, and reintroduce them into the body in greater numbers.
- Isolate T cells, genetically modify them to detect cancerous cells, and then reintroduce a large number of them.
Both of these strategies, which rely on large, highly purified cell samples, can assist an individual’s immune system in fighting specific types of cancer.
CTC Enrichment for Cancer Research
Circulating tumor cells are cells that break off from a cancerous tumor and float through the bloodstream (CTCs). These cells can be isolated and studied in a lab to learn how cancer cells respond to various treatments or environments. Obtaining highly concentrated CTC samples enables non-invasive cancer research that evaluates potential outcomes without endangering patients.
Cell Sample Purification for Protein Therapy
Protein therapy is another type of cell engineering. Protein therapy entails replacing, replenishing, or reprogramming specific cells to produce certain proteins. When a person’s cells are damaged or incomplete, scientists can repair or replace proteins to repair the damaged cell.
COVID-19 Immune Response Study
T cell isolation enables researchers to conduct a wide range of infectious disease tests. The ability to study cells involved in the SARS-CoV-2 virus and COVID-19 disease can provide clues about how to combat them. Infected cells are less abundant and necessitate gentle, precise cell separation methods in order to extract large volumes of viable cells.
Choosing the Best Cell Separation Method
The cell enrichment technique you should use is heavily dependent on your situation. If you work for a large organization or laboratory with extensive funding and stringent cell sorting requirements, it may be acceptable to invest more time and money in a more complex set of machinery.
Some cell populations can only be separated using certain methods, while others are easier to separate using one method over another. When it comes to preserving cell viability for downstream applications, doing research on the best product for your specific needs can save you a lot of trouble.
When it comes to the most cost-effective and time-efficient method for single-cell type isolations, the cheapest and quickest method that maintains cell health with high throughput is Pluriselect’s products.
Whether you are looking to further purify your sample after using another method or perform simple cell separation procedures in the most efficient way, Pluribead is the best option for speed, ease, and maintaining cell health and physiology. Our products have already helped a multitude of research efforts.
Check out our huge range of products or contact us to find out more about how our products can benefit your cell separation efforts.
Reference:
Science Direct
Nature
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