Tumor Enzymatic Digestion: A Procedure to Generate Tumor Cell Suspension from Patient-derived Xenograft Cancer Mouse Models

- Patient-derived xenograft, or PDX, models are experimental animal models bearing tumor cells or tumor fragments implanted directly from a human patient. These models simulate the human tumor tissue microenvironment, allowing tumor propagation. To generate tumor cell suspensions, first, harvest the patient-derived tumor portion from the recipient mouse.

Mince the tumor segments to obtain smaller fragments. Treat the fragments with a digest solution containing a cocktail of collagenase, hyaluronidase, and deoxyribonuclease enzymes. Collagenase and hyaluronidase enzymes digest the extracellular matrix that holds the tissue together, facilitating the release of individual cells into the suspension.

The deoxyribonuclease degrades any free DNA released from lysed cells. Now, filter the digested sample through cell strainers to remove any undigested tissue and debris and obtain single-cell suspensions. Centrifuge to separate the cells. Discard the supernatant containing the digestion mix and resuspend the cell pellet in fresh media.

Finally, supplement with lymph node stromal cells in the desired ratio to enhance the metastatic potential of the tumor cells. In the following protocol, we