A Simple and Rapid Protocol to Non-enzymatically Dissociate Fresh XXXX Tissues for the Analysis of Infiltrating Lymphocytes.

@entity5 @entity1 @entity1195 @entity0
The ability of malignant cells to evade the immune system, characterized by @entity5 escape from both innate and adaptive immune responses, is now accepted as an important hallmark of @entity5 . Our research on @entity0 focuses on the active role that @entity5 infiltrating lymphocytes play in @entity5 progression and @entity1 outcome. Toward this goal, we developed a methodology for the rapid isolation of intact lymphoid cells from normal and abnormal tissues in an effort to evaluate them proximate to their native state. Homogenates prepared using a mechanical dissociator show both increased viability and cell recovery while preserving surface receptor expression compared to enzyme-digested tissues. Furthermore, enzymatic digestion of the remaining insoluble material did not recover additional @entity1195 (+) cells indicating that quantitative and qualitative measurements in the primary homogenate likely genuinely reflect infiltrating subpopulations in the tissue fragment. The lymphoid cells in these homogenates can be easily characterized using immunological (phenotype, proliferation, etc.) or molecular (DNA, RNA and/or protein) approaches. @entity1195 (+) cells can also be used for subpopulation purification, in vitro expansion or cryopreservation. An additional benefit of this approach is that the primary tissue supernatant from the homogenates can be used to characterize and compare cytokines, chemokines, immunoglobulins and antigens present in normal and malignant tissues. This protocol functions extremely well for @entity1 breast tissues and should be applicable to a wide variety of normal and abnormal tissues.
