Cell Disruption
Cell disruption is a common technique in various analyses of cultured cells. Regardless of your desired analyte (DNA, RNA, proteins, metabolites, etc.) there are a number of common considerations for cell disruption applications.
Tips for Homogenizing Cells
Plant cells or cells that form spores are often more difficult to lyse than larger cells without cell walls, and may therefore take longer to homogenize. Cultured animal cells are generally very easy to disrupt.
Unless you have a temperature-sensitive analyte or specialized application which requires you to be gentle, turn the speed / power of your homogenizer up! It will make the homogenization faster and more efficient, and for most applications there’s no such thing as homogenizing your sample too well.
Choosing a Homogenizer for Cell Disruption
Just about all homogenizers are acceptable for cell disruption applications.
Many high-pressure homogenizers, such as the French pressure cell press, were specially designed for cell disruption. Although they can be very expensive, they are extremely efficient at homogenizing large volumes of cells. Ultrasonic homogenizers are also highly efficient at cell lysis, although they are not suitable for all applications due to the amount of heat generated. Bead mills are very capable for smaller volumes of cells. For efficient homogenization, ensure that a very small bead size is used. Rotor-stator homogenizers are not as efficient as ultrasonics at cell lysis, but they will still get the job done without creating as much heat, albeit a bit slower.
Total RNA extracted from Streptomyces. Lane 1: extracted using a Precellys® 24. Lane 2: extracted with enzymatic lysis.
If you have a low-throughput application and cost is a significant concern, a mortar and pestle can be used as well. A motorized mortar and / or a tightly fitted mortar and pestle are recommended in order to generate additional shear and thereby more effectively lyse the cells.
