The Core will provide necessary support for the creation, selection, characterization, production, and purification of recombinant adenoviruses, as well as the isolation and culture of neonatal rat cardiomyocytes and neonatal and adult mouse cardiomyocytes. The Core is currently used by all of the Projects. The Core facility is and will be directed by Christopher Baines, Ph.D., who was appointed to the faculty in the Department of Pediatrics in 2005, Division of Molecular Cardiovascular Biology. Dr. Baines has extensive experience in cardiomyocyte preparation and in the generation and use of recombinant adenoviral vectors in the study of myocardial cell biology (Baines et al, 2005). He assumed oversight of Core operations in 2005 when Dr. Orlando Bueno left. The necessary personnel are already working in the laboratory and are fully trained. Core C will provide a variety of support services including: a. Production of the shuttle vector plasmid DNA (for distribution to investigators) b. Electroporation of transgene shuttle vector into adenoviral genomic DNA-containing bacteria for recombination c. Identification and purification of positive adenoviral recombinants d. Maintenance of human embryonic kidney (HEK) 293 cells permissive for adenoviral replication e. Amplification of recombinant cultures f. Purification and concentration of recombinant adenoviruses g. Titration of stocks for determination of plaque forming unit (pfu) levels h. Repository service for storage and cataloging of low passage viral stocks to serve as a backup for viral stocks given to investigators i. Weekly preparation of neonatal rat cardiomyocyte cultures j. Preparation of neonatal and adult mouse cardiomyocyte cultures when required
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