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The overall goal of the Cell Separation and Sample Preparation (CSSP) Core is to develop technologies for the collection and isolation of enriched blood leukocyte cell populations that are applicable to critically ill patient populations for subsequent high throughput proteomic and genomic analyses, as well as for functional proteomics. The Core also assists with the collection, processing and archival of solid tissue samples obtained at surgical interventions in the trauma and burn patient populations.
Because of these distinct functions, the Core has both development and service components similar to the Genomics Core.
The challenge to the CSSP Core is to successfully balance the development of protocols that yield both a high quality, enriched leukocyte product and at the same time, are feasible in a critically ill patient population under adverse conditions. The Program investigators realize that technologies exist that can generate essentially pure cell populations for proteomic and genomic analyses, but because of blood volume, time, cost, training, equipment and other feasibility considerations, could not be applied to a multi-center clinical study. Therefore, protocols must be developed that are sufficiently robust and automated to yield highly enriched cell populations from small blood collection volumes that can be performed by trained nursing or research staff, within the time and effort constraints associated with a critically ill patient population.
The keys to success during the first five years of the Program were:
During this time, macroscale approaches for the collection and isolation of leukocytes and leukocyte subpopulations from whole blood for genomic analyses have been developed and implemented within the Program. As the Program matured, it became apparent that to obtain meaningful biological information from the genomics and proteomics data, ever increasingly enriched cell populations would be required. It is our belief that microfluidics represents the future direction for the isolation of these enriched populations.
Simultaneous with the implementation of these macroscale approaches to the collection and isolation of leukocytes and leukocyte subpopulations, the Program has developed a highly productive microfluidics approach to cell separation and nucleic acid recovery, whose primary goal over the next funding period is their implementation at the clinical sites.
The program has three microfluidics modules in varying stages of development or implementation:
Major Functions of this Core
As the Program moves forward in Years six to ten, the primary goals of the CSSP Core are:
These enriched cell populations will then be used for both proteomic (high throughput proteomic determination of cellular proteins, and functional phenotypic characterization of the cell populations) and genomic analyses.
This Core consists of two centers of excellence at the University of Florida College of Medicine and Massachusetts General Hospital/Harvard Medical School. Dr. Lyle Moldawer of UFL is the Director of the CSSP Core.
Laboratory of Inflammation Biology and Surgical Science at the Department of Surgery, University of Florida School of Medicine under the leadership of Lyle Moldawer, PhD (administrative oversight for sample collection, archival and distribution to the analytical sites, and quality assurance/control for sample collection and processing)