Cell Science Core
The Cell Science Core (CSC) is critical to the development of projects within the Center for Pediatric Research. The Core brings together all of the cell and proteomic resources available within the department and provides access to additional services available at the University of Delaware, the Delaware Biotechnology Institute, Thomas Jefferson University and Christiana Care Health Systems. The Cell Science Core provides services and resources for preparative and analytical studies for cell biology, and protein biochemistry. Additional outreach activities are the provision of resources to help the growth of local biotech companies and bring new technologies from local equipment manufactures to advance our research.
• Preparative Facility: This facility provides a centralized staffed tissue culture facility for preparation of anonymized EBV-immortalized lymphoblastoid cell lines and fibroblast cultures from human subjects, and is also involved in maintaining other immortalized cell lines to serve as a bank for these cell-based reagents.
• Analytical Facility: This facility consists of the Cell Biology Resource, the Protein Biochemistry Resource and a chromatographic separations unit.
• Proteomics Sub-Core:
This facility has been established in collaboration with the Delaware Biotechnology Institute. The facility provides the ability to determine protein expression differences using fluorescent dyes to stain different populations of proteins that can be analyzed together in a series of 2D gels by DIGE analysis. Individual proteins are picked from the gels, digested with trypsin and spotted onto MALDI plates for identification by MALDI MS/MS. Quantitative approaches to protein analysis are emphasized. Core staff personnel provide consultation for grant applications and assistance with all protein analytical techniques. MALDI MS/MS facilities are available at the INBRE-supported Delaware Biotechnology Institute and procedures have been developed to integrate the Delaware Core facilities to optimize use of this equipment. Shot-gun proteomics by LC-MS/MS are also available through the Delaware Biotechnology Institute. Three COBRE target investigators and several affiliated faculty currently use this facility. This core has strengthened ties with the University of Delaware through the Delaware Health Sciences Alliance.
• FACS Sub-Core:
The FACS sub-core continues to work closely with the Delaware INBRE to provide cellsorting for our investigators at Christiana Care by reciprocal agreements. The Accuri Flow Cytometer in the core is used extensively and has dramatically expanded the utilization of this core. It is enabling development of new projects in cancer research and provides fee-for-service for local biotech companies. Junior investigators are mentored in the use of FACS analysis and are now incorporating this technology into their research. FACS utilization continues to expand, now including routine analysis or tumor engraftment in a mouse model of leukemia.
• LC/MS/MS Sub-Core:
In late 2013 an Agilent 6460 triple quad MS/MS instrument attached to an 1100 LC instrument via an ESI interface was added to the lab. The equipment was obtained to fully evaluate the utility of this instrument for a range of studies related to drug development and clinical diagnostics for mucopolysaccharidosis diseases. The instrument will help the core move towards self-sufficiency by incorporating clinical diagnostics that will generate revenue.