Home/ Technology/ Human Monocytes/ Human Monocyte Isolation With Akadeum’s Microbubbles
Each cell in the human body has a specific role to carry out that is essential to the organism. When it comes to defending the body against foreign pathogens, there is a complex system of immune cells cooperating to eradicate harmful substances to keep the body at peak health. Isolated populations of immune cells are studied to gain knowledge of their structures, life cycles, and functions. One type of white blood cell that can be isolated from human blood samples is the monocyte.
Monocytes, the largest of the white blood cells, develop in the bone marrow before entering the bloodstream where they represent 3-8% of circulating blood cells. From the bloodstream, monocytes will migrate into tissues where they can further differentiate into macrophages or dendritic cells. Dendritic cells are mostly found in barrier tissues that come into contact with the outside environment including the skin, lungs, lining of the nose, and intestines.
Although every aspect of how monocytes function is not completely understood, it is known that they are involved in the regulation of inflammation. Dendritic cells and macrophages do this in different ways.
Dendritic cells are antigen-presenting cells (APCs) that help to activate effector T cells during an immune response. Dendritic cells can take antigens from a pathogen and use them to stimulate helper and cytotoxic (killer) T cells against the invader. Without dendritic cells, the immune system would have a difficult time identifying and eradicating foreign bodies.
Macrophages are large phagocytic cells that perform a process called phagocytosis. Phagocytosis is the process by which one cell completely engulfs and disposes of another cell or large particle. Macrophages devour damaged, dead, or dying cells in the bloodstream and tissues which are unable to carry out their normal function. One example of dead cells that need to be removed is the effector T cells remaining after an infection is cleared. These cells undergo the process of contraction, whereby most of the responding effector T cells die because they are no longer needed. Macrophages will then clear these dead and dying cells via phagocytosis.
Human monocytes are an integral part of the immune system and therefore must be studied to develop a comprehensive understanding of how humans fend off infectious diseases. To study monocytes, they must first be isolated. Human blood monocyte isolation is usually performed on peripheral blood mononuclear cell (PBMC) samples or leukopaks. Both PBMC samples and leukopaks are concentrated samples of leukocytes and other cells extracted from whole blood. The increased abundance of target cells in these samples makes it easier to enrich a sufficient number of viable cells. Purchasing PBMCs or leukopaks streamlines the cell isolation process eliminating the need to begin with freshly isolated human whole blood.
Separating target cells from a leukopak or PBMC sample can be done in a variety of ways, some more practical than others. Methods include antibody-antigen binding, sorting based on size orphysical properties, the optimal strategy is dependent on downstream use of the isolated monocytes.
Akadeum’s Human CD14+ Monocyte Cell Isolation Kit is developed specifically to isolate untouched CD14+ monocytes, using a negative selection method to remove unwanted cell populations and effectively enrich CD14+ monocytes ready for downstream use. The BACS Microbubble workflow is exceptionally gentle on cells, helping to maintain the health and physiology of monocytes during enrichment.
Additionally, Akadeum’s Streptavidin Microbubbles can target any cell marker with the proper biotinylated antibodies. The customization of our streptavidin microbubbles reduces the limitations that exist with some other separation methods. This allows scientists to benefit from microbubbles regardless of the target cell population and reduces the limitations that exist with some other separation methods.
On top of the flexibility, the process itself requires almost no additional training as it is a straightforward protocol. To use the microbubble kit, the antibody cocktail is added to the sample, a brief incubation is necessary and then microbubbles are mixed with the antibody and cell sample. This can be performed directly in the sample container and does not require additional equipment. After the microbubbles bind to labeled cells and float them to the top of the sample, the cells of interest are untouched at the bottom of the tube. Cell separation is so easy—it floats!
The entire process of using most of Akadeum’s kits takes only about 30 minutes including setup and cleanup. Our products are designed for efficiency and scalability. The kits work fast and there are no limitations as to how many samples you can process at once. If you have two samples that must be enriched, instead of waiting for one sample to run through a magnetic isolation system or flow cytometer before running the other sample, you can simply run them simultaneously. Akadeum’s microbubbles are fast and simple, but how well do they work to preserve cell health?
If you’re attempting to enrich a sample of human monocytes for downstream research or applications, you should be looking for a monocyte isolation technique with high viability. BACSTM technology uses the power of buoyant microbubbles to gently float labeled cells to the top of a sample, meaning there are no harsh forces throughout the process. Additionally, when the unwanted cells are removed from the sample the desired cells are left untouched at the bottom of the tube, as healthy as the moment they were placed into it.
Our microbubbles emphasize preserving the behavior and physiology of desired cells to ensure accurate downstream results.
Akadeum is currently working to expand our human primary cell isolation products and develop mutually beneficial partnerships along the way. We hope that by offering a solution to many of the long-standing problems in cell separation, we can help you and other fellow researchers streamline cell research assays to deliver better results.
If you think our technology could be used to optimize your human monocyte isolation efforts, reach out to one of our scientists to set up an appointment. We look forward to hearing about you, your business, and any goals that we can help you to achieve.