Proof of Concept to Isolate and Culture Primary Muscle Cells from Northern Elephant Seals to Study the Mechanisms that Maintain Aerobic Metabolism Under the Hypoxic Conditions of Breath-hold Diving

• Main Author: Kanatous, Shane B
• Other Authors: COLORADO STATE UNIV FORT COLLINS DEPT OF BIOLOGY
• Format: Text
• Language: English
• Published: 2013
• Subjects: Biology; Life Support Systems; *METABOLISM; *RESPIRATION; *SEALS(MAMMALS); AEROBIC PROCESSES; BIOPSY; CULTURE; HYPOXIA; MUSCLES; Elephant Seals
• Online Access: http://www.dtic.mil/docs/citations/ADA597966; http://oai.dtic.mil/oai/oai?&verb=getRecord&metadataPrefix=html&identifier=ADA597966

This work will be carried out in collaboration with Drs Daniel Costa, and Daniel Crocker. We will collect muscle biopsies from animals that are being sedated for their labs respective projects. By utilizing the same animals and working with these collaborators, we will minize the impact and number of animals being handled at A o Nuevo State Reserve. Muscle sampling Biopsy samples of approximately 50 mg will be collected with a 6-mm biopsy cannula (Depuy, Warsaw, Indiana) from the swimming (M. longissimus dorsi) muscle. Before the biopsy is taken, the skin is first cleaned with betadine scrub. A small area the size of a dime is shaved and one ml of 2% Lidocaine is then injected beneath the skin around the biopsy site to minimize the trauma to the biopsy site. The skin is punctured with a #10 scalpel and the biopsy needle inserted to a depth of 3-4 inches (the blubber layer is typically 2 inches thick). Once collected, the biopsy will be dipped in 100% ethanol for sterilization then placed into culture media (F10 or F10 plus lipids) and stored on ice. After collection the biopsy site will be cleaned with betadine scrub. Slight pressure is applied to the biopsy site after flushing with 4x4 gauze pads to stop any minor bleeding. The biopsy samples will then be shipped overnight on ice to the lab at Colorado State University for primary cell isolation and culturing.