One of the applications we are asked about most frequently is using a homogenizer for tissue dissociation - isolating intact cells from tissue. Can it be done? Yes, it most certainly can, but it is a finicky process. Here's some advice for using a homogenizer in cell isolation applications:
- Don't use ultrasonic homogenizers for cell isolation. Ultrasonic homogenizers are equally effective, if not more effective, at lysing cells as they are at dissociating tissue. If you manage to break apart your tissue on a macro-level you'll probably have lysed your cells as well.
- It's possible to perform cell isolation using a rotor-stator if you can run it very slowly and / or use a blending / cutting attachment. Some rotor-stators have minimum speeds which prevent them from being run slow enough to shred tissue without lysing cells due to the shear forces it creates. However, some rotor-stators have attachments which spin a blade, turning the device into more of a blender than a traditional rotor-stator. These generally improve tissue separation while limiting cell lysis due to shear forces.
- Bead mill homogenizers are usually our recommendation for tissue dissociation (although it can depend on other parameters as well). As with rotor-stators, you'll want to minimize your run speed and run for only as long as you need to dissociate the tissue. This generally takes some experimenting to find your ideal run time and speed which will gently dissociate the tissue. Using larger beads than would normally be used is recommended, as the collisions between beads (or collisions between the beads and the wall of the tubes) tend to increase the efficiency of cell lysis, which is undesirable in this case. Note that your cell population may have slight contamination from micro-particles that chip off the beads. In most research applications this mild contamination doesn't matter, but for some preclinical applications or certain downstream analytical measurements it may cause a problem.
Ultimately, homogenizers are designed for homogenizing. If you want to use one for cell isolation you're not going to get a very high percentage of viable cells and you'll probably have to do some experimenting to perfect your protocol, but ultimately it is possible and isn't terribly difficult to do with most tissues.
If you're looking for some assistance using a homogenizer for cell isolation or any other application, feel free to contact us!