Proteins are indispensable tools for science research and the study of living organisms. Because they are essential to both cell structure and function, much can be learned through protein analysis; these molecules can be accessed through cell rupture, which entails disrupting the cell wall and/or membrane.
In particular, mammalian cell lines are more commonly used than any other cell type because of their easy maintenance, scalability, and genetic modifiability.
Below is a compilation of factors, specific to mammalian cells, that should be considered as you optimize your cell rupture technique.
1. Chosen Technique
Unlike most other cell types with tough exterior walls, the contents of mammalian cells are only separated from the extracellular environment by a thin membrane. Multiple cell rupture techniques exist, such as homogenization, mechanical shearing, freezing, and sonication.
Because primary and culture mammalian cells do not require physical disruption, detergent-based cell lysis is an easy alternative that can be combined with homogenization for optimal results. Detergents work to break apart the lipid bilayer, and homogenizers finalize particle breakage, so that all contents of interest can be accessed.
2. Protein Location & Stabilization
Both cellular location and chemical stability play major roles in determining the best way to approach cell rupture. For example, certain proteins are located in specific organelles. Rupture methods that release the contents from every compartment make it incredibly difficult to achieve high yield of a specific protein of interest.
In contrast, optimization of methods that are able to release contents from specific structures makes the process easier, and enhances product isolation.
3. Tissue Type
Mammalian organisms are made up of four major tissue types: connective, epithelial, and nerve & muscle. These can be further differentiated into specialized tissues, all of which have unique structures. The tissue type your cell hails from should impact your cell rupture method, as the method may differ with tissues.
According to the Thermo Scientific Cell and Protein Isolation Technical Handbook, physical disruption, such as homogenization, is the best option for cells within tissues, as they are bound tightly within organ-specific matrices. In contrast, primary and cultured mammalian cells simply require some form of enzyme exposure or mechanical dissociation.
Pion: The Homogenizer Advantage
Whether working with mammalian, yeast, bacterial, or viral cells, it is important to consider the equipment you are using for your cell rupture technique. As the preferred method, high pressure homogenization will save precious time while still achieving a high yield of intracellular contents.
Pion's BEE brand technology manufactures high pressure homogenizers that are trusted by researchers and lab managers around the world. We deliver an array of key benefits- importantly, cell rupture- but also production of nano/micro emulsions and dispersions and lipids and suspensions. These can be used for products across the pharmaceutical, biotechnology, food, and chemical industries.