Modeling Ovarian Cancer Cell Colonization of Omentum: An Ex Vivo Method to Establish Milky Spot Colonization of Fluorescently Labeled Ovarian Cancer Cells in Murine Omentum Explants

Abstract

Source: Krishnan, V. et al. In Vivo and Ex Vivo Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose. J. Vis. Exp. (2015)

In this video, we establish an ex vivo omental organ culture overlaid with GFP-expressing ovarian cancer cells to study ovarian cancer colonization within the omental explant. This protocol analyzes immune cell structures within the omentum known as milky spots that promote metastases.

Protocol

All procedures involving animals have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Milky Spot Colonization Ex Vivo

1. Establishment of basic ex vivo omental organ culture conditions
   1. Place each omentum into a single culture plate insert and place within one well of a 12-well tissue culture plate containing 500 µl of DME/F12 media containing 20% FCS. Maintain organ cultures at 37 °C in a 5% CO₂ environment for 24 to 48 hr and/or additional time points. Use three independent omenta (triplicate samples) for each condition (e....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Dissection Tools	Fine Science Tools	NA
Giemsa	Fluka/Sigma Aldrich	48900
5% Formalin	Sigma	HT501320
DMEM	Corning	10-013-CV
PBS	Corning	21-040-CV	Without calcium and Magnesium
Collagenase	Worthington	LS004196
Millicell culture plate insert	Millipore	PICM01250
Cell-Tak	Corning	354240