Graduate Student
Department of Microbiology and Immunology
Temple University
Experiments that involve primary cells often require the use of some mechanistic or enzymatic means to acquire a single cell suspension. Experimental variation due to differences in the acquisition of single cell suspensions is a common concern. Miltenyi Biotec has developed a solution, the gentleMACS™ Dissociator. The gentleMACS™ Dissociator is a mechanistic approach to disrupting tissue into single cell suspensions or a homogenate. The specially adapted tubes that work with the dissociator act as an organ blender of sorts. The gentleMACS™ Dissociator does what my hands and a plunger cannot accomplish as well, as quickly, or as consistently. Moreover, the gentleMACS™ Dissociator has programs specific for certain cell types; Miltenyi also has developed and posted online protocols for specific cell types. The gentleMACS™ Dissociator can handle two tubes/samples at a time.
When I began using the gentleMACS™ Dissociator I was needed single cell suspensions from mouse intestines for flow cytometry and magnetic cell isolation. While there was no posted protocol for this type of tissue dissociation, my Miltenyi sales representative put me in contact with a staff scientist at Miltenyi who sent an unpublished protocol from another investigator that was using the gentleMACS™ Dissociator to dissociate intestinal tissue. I adapted the protocol for my experiments and compared the gentleMACS™ Dissociator to the previous methods I had used to dissociate tissue. Compared to enzymatic digestion alone, the gentleMACS™ Dissociator not only more effectively dissiociated the tissue, the amount of cellular debris and cell death was reduced. I observed a more consistent and through digestion of intestinal tissue, even with increased amounts of sample tissue in the digestion buffer. Cells were easily run through a cell strainer and prepared for density gradient separation.
The gentleMACS™ Dissociator does have quite a bit of upfront cost. However, if your experiments involve difficult tissues, low cell number, or high sample numbers the gentleMACS™ Dissociator may prove to be a time saver and thus, worth the cost. The tubes are an ongoing cost; however, if your experiments do not require absolute sterility they can be washed and used again.