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Acinar-to-Ductal Metaplasia Induction: A Method to Study Acinar Cell to Ductal Cell Differentiation in 3D Ex Vivo Culture
Overview
This video describes the technique of differentiating pancreatic acinar cells to ductal cells in ex vivo 3D culture. This helps to understand the mechanisms regulating the acinar-to-ductal metaplasia (ADM), a key process in the early development of pancreatic cancer.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Preparation of Materials, Solutions, and 3-dimensional Matrix Bases
- Cut 500 µm and 105 µm polypropylene meshes into 76 mm by 76 mm squares. Fold each square in half twice to create a smaller folded square. Place one 500 µm and one 105 µm mesh square into an autoclavable pouch.
- Autoclave polypropylene mesh s.......
Representative Results
Figure 1: Representative results of primary acinar cells plated within collagen after stimulation with TGF-α. Primary acinar cells from a non-transgenic mouse were infected with GFP-adenovirus, embedded in collagen I or basement membrane matrix, and stimulated with or without TGF-α (50 ng/mL). Twenty-four hours post-infection with GFP adenovirus, images were captur.......
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Source: Fleming Martinez, A. K. et. al. Mimicking and Manipulating Pancreatic Acinar-to-Ductal Metaplasia in 3-dimensional Cell Culture. J. Vis. Exp. (2019).
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