This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. During the past three years there was a noticeable increase in the use of the cell culture facility with the consequential crowding and scheduling conflicts. Most of the users use nerve cells, either neurons, glia or cell lines that are models of neural cells. The multiplicity of users increases costs, crowding and expense. By dedicating one technician for most of the preparations will alleviate most of those problems. The proposed core facility will facilitate the UCC investigators the use of cultured neurons, glias and organotypic cultures. The investigators will no longer need to develop by themselves the techniques to culture brain cells or brain tissue (organotypic cultures). The cultures will be used, among others, for patch clamping after in vitro treatments with drugs of addiction or inhibitors and for in vitro models of neurodegenerative diseases. The core facility will consist of a coordinator (PAF) and a technician (Brenda Cuadrado) who already has three years of experience in neuronal cell culture. Among other skills she is able to prepare neuronal cultures from fetal cortex or hippocampus or astrocytes from cerebral cortex. She has experience in culturing organotypic hippocampal slices on translucent and permeable plastic membranes using the Stoppini method. Mrs. Cuadrado was trained in the UCC (by PAF), by Dr. C. Ghiani from UCLA and she traveled to Lexington, Kentucky University, to learn organotypic slices culture in the laboratory of Dr. Littletone. The growth and continuous up grading of our services will be facilitated thanks to our close collaboration with Dr. J. de Vellis, who is an expert in glia and neuron culture. Dr. J. de Vellis recommended as a direct consultant Dr. Steve Levison. The model of the proposed service unit is based on the arrangement of Dr. de Vellis? laboratory (UCLA) with a single person dedicated to provide glial and neuronal preparations to this prestigious group. The proposed budget is mainly the salaries of the technician (Mrs. Cuadrado), consultant and the coordinator. Funds are requested for starting material and supplies to allow for a smooth transition to a system of fee for service. New equipment is requested to modernize and replace more than10 years old incubators and so prevent problems in the near future. The objective of this unit is not to exclude anyone from using the existing cell culture facility but by providing a better alternative discourage the initiation of new neuronal culture laboratories throughout the UCC. By doing so we will increase the efficient use of space and equipment. By providing the expertise in brain cell culture and the labor involved this core facility will leave more time to the investigators for the scientific endeavor.
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