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. The class is offered to individuals involved in collaborative projects. The training focuses on the hands-on manipulation of all essential skills to allow the individual to carry out experiments that involve the use of cultures cells and/or engineered tissues. Current training includes: Basic sterile cell culture techniques, methods of cell preservation and recovery, cell counting, culture of cells in Rose Chambers, dissociation of human tissues of normal dermal and pathological dermal in origin and human glioblastoma to establish cell lines for future use, tumor spheroid and raft culture in collagen and fibrin. In addition, we provide training in Shellless Chick Embryo CAM (Chorioallantoic Membrane) model with tumor spheroids implantation. TUNEL assay, actin visualization and various other labeling methods that allows the MPM alone or MPM/OCT imaging of specific subcellular organelles and intermediate filaments with the purpose of monitoring the consequence of therapeutically treatment and dynamics of cell function in extracellular matrix. In the past year we have developed our training classes. We started tumor stem cells isolation with special focus on human glioblastoma CD133 positive cells with the first human glioblastoma biopsy received in mid-December 2006 (work in progress). Digital images of the procedure to establish Shellless Chick Embryo CAM models were acquired and used for training. These images were widely requested by parties trained from other institutions. Our complete Raft culture protocol has also been widely requested. We have received many requests and have provided training for establishment of tumor spheroids. In the coming year we will focus on posting all existing protocols on-line with digital images attached for access by off site collaborators with password access and for in house self guided training.
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