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Pancreatic Islets Isolation through Collagenase Perfusion: An Enzymatic Method to Isolate Pancreatic Islet Cells
Overview
Pancreatic islets, also called the Islets of Langerhans, are a cluster of endocrine cells which produces hormones for glucose regulation and other important biological functions. This protocol explains a method for isolating pancreatic islets by first perfusing collagenase via Ampulla of Vater to digest the exocrine pancreatic tissue and a simplified density gradient centrifugation to purify the islets from mice.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
The surgical tools needed are shown in Figure 1 and the schematic diagram of the procedure is shown in Figure 2.
1. Solutions
- Prepare Hank’s Balanced Salt Solution (HBSS) by adding 100 mL of 10X HBSS (from stock) to 900 mL of distilled water to make 1 L HBSS (1X).<.......
Representative Results
Figure 1: Surgical tools.
Curved surgical scissors, cover glass forceps, micro Adson forceps, curved forceps, small surgical scissors, and Schwartz micro serrefines (microvascular clamp) are shown. Please click here to view a larger version of this figure.
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Source: Villarreal D. et al. A Simple High Efficiency Protocol for Pancreatic Islet Isolation from Mice. J. Vis. Exp. (2019)
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