Modeling Ovarian Cancer Communication for Imaging Mass Spectrometry Analysis: A Method to Identify Small Molecule Chemical Communicators in Metastatic Ovarian Tumor Development

Abstract

Source: Zink, K. E. et. al. Capturing Small Molecule Communication Between Tissues and Cells Using Imaging Mass Spectrometry. J. Vis. Exp. (2019)

This protocol describes a technique for sample preparation by co-culturing ovarian explants with the fallopian tube cancer cells. These samples are visualized using imaging mass spectroscopy, or IMS,  to detect small molecule exchange between the cells and tissues leading to tumor development.

Protocol

All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Setting Up and Incubating the ITO-treated Slide for Cocultures Undivided cocultures Liquify the agarose at 70 °C on a hot plate. Place the 8-well divider on top of the indium tinoxide (ITO)-treated slide (Figure 1A). The rubber bottom on the divider aids in…

Representative Results

Figure 1: Workflow for sample preparation of undivided coculture. (A) Adhere 8-well chamber to conductive side of ITO-coated slide. (B) Place halved ovaries in the center of wells for coculture conditions. (C) Add 300 μL of agarose/cell suspension directly into wells. Make sure there are no air bubbles and that the ovary rema…

Materials

Denville	C1017-O	To collect cells
Millipore	PEZGS0816	Repurposed from Millipore Millicell EZ-slide chamber slide
Sigma-Aldrich	34998-4L	Solvent for sprayed matrix
Eppendorf	5810 R	To collect cells and remove supernatant
Bruker Daltonic	8201344	Matrix sprayed onto dried slide
Bruker Daltonic	8201346	Matrix sprayed onto dried slide
Gibco	11995-065	Media mixed with agarose
Genesee Scientific	22-282	For agarose aliquots
Bruker	8237001	Platform for co-culture incubation
Sigma-Aldrich	A9414-10G	Mixed with media for plating
Corning	4870	Used to cut plastic dividers for divided chambers
Bruker Daltonic	8206195	Calibrant for medium mass range
Sigma-Aldrich	343-242-5G	Calibrant for low mass range
Simga-Aldrich	E2758	Cell culture media supplement
Peprotech Inc.	100-15	Cell culture media supplement
Cellgro	30-005-CR	Cell culture media supplement
Sigma-Aldrich	11074547001	Cell culture media supplement
Gibco	25030-081	Cell culture media supplement
Gibco	15140-122	Cell culture media supplement
Fisher Technologies	A116-50	Added to matrix solution
HTX Technologies		For applying matrix