Modeling Ovarian Cancer Communication for Imaging Mass Spectrometry Analysis: A Method to Identify Small Molecule Chemical Communicators in Metastatic Ovarian Tumor Development
Published: April 30, 2023
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…
Açıklamalar
The authors have nothing to disclose.
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 |
Etiketler
Bu Makaleden Alıntı Yapın
Modeling Ovarian Cancer Communication for Imaging Mass Spectrometry Analysis: A Method to Identify Small Molecule Chemical Communicators in Metastatic Ovarian Tumor Development. J. Vis. Exp. (Pending Publication), e20372, doi: (2023).