75: Using an ex-vivo Human Tumor Platform to Perform Dynamic Live Tumor Imaging

Saturday, March 12, 2022

11:51 AM – 11:59 AM CST

Location: Chantilly West

Abstract Presenter(s)

Areeba Saif, MD

Surgical Oncology Research Fellow
National Institutes of Health
Rockville, Maryland, United States

Disclosure: I do not have any relevant financial / non-financial relationships with any proprietary interests.

Participants should be aware of the following financial/non-financial relationships:

Areeba Saif, MD: I do not have any relevant financial / non-financial relationships with any proprietary interests.

Introduction:

The ability to accurately predict how a patient’s tumor will respond to therapy will allow for the optimization of truly personalized cancer care. To this end, our lab produced a novel translational platform, the SMART System (Surgically-resected Mesothelium ContAining UnalteRed Tumor Microenvironment) that faithfully recapitulates tumor biology by preserving the innate structure, cell-cell interactions, and normal human physiology to mirror in vivo biology, albeit in an ex vivo environment amenable to detailed interrogation including advanced imaging techniques and live imaging, in particular.

Methods:

A clinically indicated diagnostic laparoscopy is performed and tumor-bearing peritoneum is procured from the patient. Tissue is affixed to specialized 3D-printed platforms and maintained in the SMART system. The system accommodates the application of drug(s) in the perfusate and subsequent interrogations by live-cell imaging. A customized 3D-printed imaging adaptor, made from autoclavable photopolymer resin and mounted on a microscope stage incubator, is used to perform imaging.

Results:

Prolonged live-cell imaging on tumor-bearing peritoneum for up to twelve hours is performed on patients with gastric cancer peritoneal carcinomatosis and mesothelioma. Fluorophore-conjugated anti-CD44 and anti-CD45 antibodies are used to highlight tumor cells and immune cells respectively. Using a stage incubator and optimized gas mixer for O2 and CO2 delivery, we demonstrate good tissue preservation and stable immune cell mobility for up to twelve hours. Tumor-infiltrating immune cells are tracked over time and both immune cell mobility and immune-tumor cell ratio are quantified prior to and post IL-2 administration. IL-2 stimulation in our system leads to enhanced immune cell mobility which is quantified using an image analysis software.

Conclusions:

We developed an ex-vivo human perfusion system that allows us to perform dynamic prolonged live-cell imaging on various tumor samples.

Learning Objectives:

Upon completion, participant with be able to describe the specialized ex-vivo perfusion model designed to characterize the tumor microenvironment of various tumors.
Upon completion, participant will be able to demonstrate an understanding of live tumor imaging using different cell surface markers for immune and tumor cells.
Upon completion, participant will be able to understand changes in the tumor microenvironment in response to drugs/cytokines added to our ex-vivo perfusion system.