CORE C: Cell and Molecular Biology - Ying Jin, Ph.D. - PI This Core has existed for 10 years within the Department of Neurobiology and Anatomy to facilitate the molecular analysis of cells and tissues in the SCI site and within transplants. Most importantly, the core prepares cells for grafting experiments in the individual projects. All projects will use the Cell and Molecular Core. The facility isolates, expands and stores primary fibroblasts, neural stem cells and cell lines as required. We genetically modify cells by viral transduction or transfection and provide quality control for all cells and reagents used in the core. The staff of the Cell and Molecular Core provides assistance for experimental design and data analysis. The core has developed a computerized ordering system in which individual investigators &staff request control, labeled or genetically modified cells for transplantation. Our records show that over the past 5 years the core has provided over 3 x 1 0 ^ cells, including unmodified and modified fibroblasts, neuronal and glial restricted precursors (NRP and GRP), neuroepithelial cells (NEP) and cells from immortalized cell lines. The core maintains viruses and transduced cell stocks in long-term storage and serves as a database and repository for cDNA clones, vectors, viruses and cells. Efficient and cost-effective ordering of tissue culture and molecular biology supplies as well as large scale testing of new sera and improved growth media, enzymes, kits or equipment needed for molecular analysis (thermocyclers, Q-PCR) are key core functions. We are also continuously developing new improved plasmids and viruses and have recently added and adeno-associated virus vector (AAV) as an option to our program. This will allow us to specifically transfect graft neural stem cells with growth promoting and therapeutic factors to be delivered at or near the site of injury.
The complexity of the proposed projects using genetically modified cells for transplantation and delivery of therapeutic genes requires a central facility with technical expertise, equipment and ongoing training. The core provides uniform populations of cells with minimal variations and molecular analysis to allow for inore direct comparisons betiA/een different injury models and treatment strategies.
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|Ketschek, A R; Haas, C; Gallo, G et al. (2012) The roles of neuronal and glial precursors in overcoming chondroitin sulfate proteoglycan inhibition. Exp Neurol 235:627-37|
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