IACUC / LARC STANDARD PROCEDURES

ISLET TRANSPLANTATION UNDER THE KIDNEY CAPSULE IN MICE (Posted May 2007)

Description of procedure:

Islet Transplantation (following the NIH recommended procedure): Islets are obtained from the pancreas of euthanized donor mice that have been cervically dislocated. Donor mice will not be euthanized with CO2 prior to cervical dislocation because of the change in blood and tissue pH due to the CO2 exposure. It has been shown that islet beta cell function and enzymes used to locate the islet beta cells are sensitive to pH changes. Only personnel who have sufficient experience with conscious mice will carry out the cervical dislocation of the donor mice. Mice will be firmly restrained and quickly euthanized. A mid-line incision is then made and the diaphragm is cut. The donor pancreas can then be removed for isolation of the beta islets.

The recipient mice are anesthetized and pre-operative analgesics are administered. The surgical area of the mouse will be shaved and cleaned with iodine or chlorhexidine solution and aseptic techniques practiced. A small incision is made in the left flank and the left kidney is exposed. Approximately 500-1000 islets are transplanted under the kidney capsule using a Hamilton syringe with P-150 polypropylene tubing. Hemostasis is maintained, the tissue is moistened with saline and the kidney is placed back into the abdominal cavity. The peritoneum is then closed with 5-0 silk sutures and the skin is closed with surgical glue or wound clips. Antibiotics may be indicated for diabetic or immuno-compromised animals. The mouse is allowed to recover in the cage with a heat lamp or heat pad, under close supervision. If surgical clips are used, they are removed 10-14 days post surgery. Transplanted mice will be followed until rejection or 100 days at which time the mice will be euthanize.

Mice will be evaluated for general appearance, ability to move normally around cage and reach food and water, ability to eat and drink, grooming habits, skin color, lethargy, aggression (animals will be housed individually if aggressive behavior is shown). Mice will be evaluated by the Body Condition scoring index. Mice will be monitored continuously after anesthesia until ambulatory. Mice are then monitored daily for the first 3-5 days and weekly thereafter. Mice with Body Condition score < 2 will be euthanized.

Literature search words required:

Literature search was performed for refinement of this Standard Procedure on April 9, 2007

Key Words	Search Site	Years Covered
Islet transplantation in rodents	Pubmed	1968-2007

Agents:

Anesthestics, antibiotics, analgesics

This procedure requires, general anesthesia (local anesthesia recommended) and systemic analgesics. Immuno-compromised animals may receive antibiotics. All agents administered to animals should be listed in the "Agents" section of RIO.

Adverse Effects:

Procedure, Agent or Phenotype	Potential Adverse Effects	Management
Kidney capsule grafting	Rejection	Euthanasia
Kidney capsule grafting	Infection	Consult LARC veterinarian

Monitoring Parameters:

Monitoring Parameters	Frequency	PI/Lab will Document
General appearance/activity level	Daily for first 3-5 days and then weekly thereafter	No
Body condition scoring	Daily for first 3-5 days and then weekly thereafter	No - unless indicated
Examine incision	Daily for first 3-5 days and then weekly thereafter	No

Describe the conditions, complications, and criteria (e.g. uncontrolled infection, loss of more than 15% body weight, etc.) that would lead to removal of an animal from the study, and describe how this will be accomplished (e.g. stopping treatment, euthanasia).

Euthanasia criteria: BCS < 2; significant surgical dehiscence or other incision problems that cannot be managed with veterinary care.

For all investigators housing animals with tumor formation, skin lesions, neurological deficits, or that are in Category E, list the expected characteristics/clinical presentations and endpoints of the animal model and the criteria for euthanasia. Note: The IACUC also requires such lists to be posted in the respective animal rooms and monitored by the IACUC compliance staff and LARC, to assure PI adherence to the endpoints listed.

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