Home › Microbiology › A Method for Murine Islet Isolation and Subcapsular Kidney Transplantation
Microbiology JoVE (Open Access) Citable · DOI

A Method for Murine Islet Isolation and Subcapsular Kidney Transplantation

DOI: 10.3791/2096-v
What you'll learn
  • ✓ Identify mouse pancreatic islets after isolation
  • ✓ Perform bile duct cannulation for pancreas perfusion
  • ✓ Isolate islets by collagenase digestion and purification
  • ✓ Transplant islets under the kidney capsule
Protocol

Biopharma Insights Transplantation of isolated islets has been proposed to be a potential treatment for type 1 diabetes. Here we describe a method to isolate islets from mouse pancreata and transplant them to the subcapsular space of the kidney.

Difficulty
advanced
Total time
~3-4 hrs per isolation and transplant
Model organism
Mouse (e.g., C57BL/6)
Biosafety
BSL-1

Steps

1
Cannulate and perfuse the bile duct

Cannulate the common bile duct and perfuse the pancreas with collagenase solution to digest the tissue.

▶ 01:49
2
Isolate pancreatic islets by digestion

Incubate the perfused pancreas in collagenase, then separate islets from exocrine tissue using density gradient centrifugation.

▶ 05:07
3
Hand-pick islets for purity

Select islets under a microscope based on size and morphology, transferring them to culture medium.

▶ 09:14
4
Transplant islets under kidney capsule

Expose the kidney and inject the isolated islets into the subcapsular space using a catheter.

▶ 12:37
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