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.
|Twiss, Jeffery L; Fainzilber, Mike (2016) Neuroproteomics: How Many Angels can be Identified in an Extract from the Head of a Pin? Mol Cell Proteomics 15:341-3|
|Detloff, Megan Ryan; Quiros-Molina, Daniel; Javia, Amy S et al. (2016) Delayed Exercise Is Ineffective at Reversing Aberrant Nociceptive Afferent Plasticity or Neuropathic Pain After Spinal Cord Injury in Rats. Neurorehabil Neural Repair 30:685-700|
|Sachdeva, Rahul; Farrell, Kaitlin; McMullen, Mary-Katharine et al. (2016) Dynamic Changes in Local Protein Synthetic Machinery in Regenerating Central Nervous System Axons after Spinal Cord Injury. Neural Plast 2016:4087254|
|Jin, Y; Bouyer, J; Shumsky, J S et al. (2016) Transplantation of neural progenitor cells in chronic spinal cord injury. Neuroscience 320:69-82|
|Sachdeva, Rahul; Theisen, Catherine C; Ninan, Vinu et al. (2016) Exercise dependent increase in axon regeneration into peripheral nerve grafts by propriospinal but not sensory neurons after spinal cord injury is associated with modulation of regeneration-associated genes. Exp Neurol 276:72-82|
|Yuan, Xiao-bing; Jin, Ying; Haas, Christopher et al. (2016) Guiding migration of transplanted glial progenitor cells in the injured spinal cord. Sci Rep 6:22576|
|Twiss, Jeffery L; Kalinski, Ashley L; Sachdeva, Rahul et al. (2016) Intra-axonal protein synthesis - a new target for neural repair? Neural Regen Res 11:1365-1367|
|Hayakawa, Kazuo; Haas, Christopher; Fischer, Itzhak (2016) Examining the properties and therapeutic potential of glial restricted precursors in spinal cord injury. Neural Regen Res 11:529-33|
|Jin, Ying; Bouyer, Julien; Haas, Christopher et al. (2015) Evaluation of the anatomical and functional consequences of repetitive mild cervical contusion using a model of spinal concussion. Exp Neurol 271:175-88|
|Hayakawa, Kazuo; Haas, Christopher; Jin, Ying et al. (2015) Glial restricted precursors maintain their permissive properties after long-term expansion but not following exposure to pro-inflammatory factors. Brain Res 1629:113-25|
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