Performing Simultaneous Euglycemic Glucose Clamp Experiments on Awake Farm Swine: A Novel Methodological Approach
Location
Philadelphia, PA
Start Date
9-5-2018 1:00 PM
Description
Background: The use of swine in biomedical and translational research is common due to their anatomic and physiologic similarity to humans. This is important when conducting PK/PD studies in which drug-dosing requirements and blood draw volumes are similar to humans. To conduct these studies effectively and minimize cost, we developed a method for 2 or more animals to be studied in slings after implantation of bilateral central venous catheter ports.
Methods: A supporting frame for the animal sling was constructed to allow the animals to remain in a non-weight bearing position. Swine were sedated with 1.0-1.5mg/dL telazol IM prior to all procedures. Under general anesthesia, central venous catheters with infusaports were implanted bilaterally in the internal jugular vein one week prior to start of experiments. CSII catheters were inserted in the animal’s abdomen. The animal was positioned in the sling apparatus, and 1mg/dL Telazol was administered IM throughout the experiment as clinically indicated.
Once sedated and in the sling, infusaport sites were cleaned with Chlorhexidine, and a Huber needle was inserted into each infusaport. The right infusaport was connected to 3 stopcocks in series for octreotide infusion, and 10% dextrose infusion. The left infusaport was attached to a VAMP Plus blood sampling system. Five team members required to run the study assumed their positions and an insulin bolus (0.15 units/kg) was administered. 30 minutes later the same procedure was repeated for the second swine. Blood draws were performed every 5 minutes. The following outlines the 5 key positions of the study team for each swine:
VAMP Operator (Blood Draws), YSI Operator (glucose analysis), Glucose Clamp Operator, Glucose Infusion Rate Monitor, Centrifuge Operator
Results: Using two swine 65 kg-85 kg, 24 glucose clamp studies were completed in a 3-week period. The use of 1mg of IM Telazol every 1-2 hours was effective in keeping the animals calm and comfortable. SpO2 and heart rate were monitored and remained within normal ranges. The infusaports were easily accessed and remained functional for the duration of the study (1,454 blood samples). The mean blood glucose was maintained between 83.7-86.4 mg/dL for the 24 experiments.
Conclusions: To our knowledge, the methodology of performing simultaneous euglycemic glucose clamp studies in large awake swine has not been described. All aspects of the study protocol were efficiently carried out and the animals remained in good health for the duration of the study. Establishing work stations with specific functions for each team member is critical for the success of these types of studies.
Embargo Period
5-30-2018
Performing Simultaneous Euglycemic Glucose Clamp Experiments on Awake Farm Swine: A Novel Methodological Approach
Philadelphia, PA
Background: The use of swine in biomedical and translational research is common due to their anatomic and physiologic similarity to humans. This is important when conducting PK/PD studies in which drug-dosing requirements and blood draw volumes are similar to humans. To conduct these studies effectively and minimize cost, we developed a method for 2 or more animals to be studied in slings after implantation of bilateral central venous catheter ports.
Methods: A supporting frame for the animal sling was constructed to allow the animals to remain in a non-weight bearing position. Swine were sedated with 1.0-1.5mg/dL telazol IM prior to all procedures. Under general anesthesia, central venous catheters with infusaports were implanted bilaterally in the internal jugular vein one week prior to start of experiments. CSII catheters were inserted in the animal’s abdomen. The animal was positioned in the sling apparatus, and 1mg/dL Telazol was administered IM throughout the experiment as clinically indicated.
Once sedated and in the sling, infusaport sites were cleaned with Chlorhexidine, and a Huber needle was inserted into each infusaport. The right infusaport was connected to 3 stopcocks in series for octreotide infusion, and 10% dextrose infusion. The left infusaport was attached to a VAMP Plus blood sampling system. Five team members required to run the study assumed their positions and an insulin bolus (0.15 units/kg) was administered. 30 minutes later the same procedure was repeated for the second swine. Blood draws were performed every 5 minutes. The following outlines the 5 key positions of the study team for each swine:
VAMP Operator (Blood Draws), YSI Operator (glucose analysis), Glucose Clamp Operator, Glucose Infusion Rate Monitor, Centrifuge Operator
Results: Using two swine 65 kg-85 kg, 24 glucose clamp studies were completed in a 3-week period. The use of 1mg of IM Telazol every 1-2 hours was effective in keeping the animals calm and comfortable. SpO2 and heart rate were monitored and remained within normal ranges. The infusaports were easily accessed and remained functional for the duration of the study (1,454 blood samples). The mean blood glucose was maintained between 83.7-86.4 mg/dL for the 24 experiments.
Conclusions: To our knowledge, the methodology of performing simultaneous euglycemic glucose clamp studies in large awake swine has not been described. All aspects of the study protocol were efficiently carried out and the animals remained in good health for the duration of the study. Establishing work stations with specific functions for each team member is critical for the success of these types of studies.