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University of Alabama at Birmingham - UAB

4 ARTICLES PUBLISHED IN JoVE

Biology

Bioenergetic Profile Experiment using C2C12 Myoblast Cells

David G. Nicholls¹, Victor M. Darley-Usmar², Min Wu³, Per Bo Jensen³, George W. Rogers³, David A. Ferrick³

¹Buck Institute for Age Research, Novato, CA, ²Department of Pathology, Center for Free Radical Biology, University of Alabama at Birmingham - UAB, ³Seahorse Bioscience, North Billerica, MA

A description of a method for profiling mitochondrial function in cells is provided. The mitochondrial profile generated provides four parameters of mitochondrial function that can be measured in one experiment: basal respiration rate, ATP-linked respiration, proton leak, and reserve capacity.

Medicine

Orthotopic Xenografting of Human Luciferase-Tagged Malignant Peripheral Nerve Sheath Tumor Cells for in vivo Testing of Candidate Therapeutic Agents

Amy N. Turk¹, Stephanie J. Byer¹, Kurt R. Zinn², Steven L. Carroll^1,3

¹Department of Pathology, University of Alabama at Birmingham - UAB, ²Department of Radiology, University of Alabama at Birmingham - UAB, ³Department of Cell Biology and Neurobiology, University of Alabama at Birmingham - UAB

A method for reliably grafting luciferase-tagged human malignant peripheral nerve sheath tumor cells into the sciatic nerve of immunodeficient mice is described. The use of bioluminescence imaging to demonstrate proper establishment of tumor grafts and criteria for random segregation of animals into study groups are also discussed.

Immunology and Infection

Bioenergetics and the Oxidative Burst: Protocols for the Isolation and Evaluation of Human Leukocytes and Platelets

Philip A. Kramer^*1, Balu K. Chacko^*1, Saranya Ravi¹, Michelle S. Johnson¹, Tanecia Mitchell¹, Victor M. Darley-Usmar^*1

¹UAB Mitochondrial Medicine Laboratory, Center for Free Radical Biology, Department of Pathology, University of Alabama at Birmingham

Blood leukocytes and platelets can be used as a marker of overall bioenergetic health of an individual and so have the potential to monitor pathological processes and the impact of treatments. Here we describe a method to isolate and measure mitochondrial function and the oxidative burst in these cells.

Cancer Research

Defining Gene Functions in Tumorigenesis by Ex vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells

Jody Fromm Longo¹, Stephanie N. Brosius^2,3, Steven L. Carroll¹

¹Department of Pathology and Laboratory Medicine, Medical University of South Carolina, ²Department of Pediatrics and Neurology, Perelman School of Medicine of the University of Pennsylvania, ³Division of Child Neurology, Children's Hospital of Philadelphia

Here, we present a protocol for performing gene knockouts that are embryonic lethal in vivo in genetically engineered mouse model-derived tumors and then assessing the effect that the knockout has on tumor growth, proliferation, survival, migration, invasion, and the transcriptome in vitro and in vivo.