ADVANCED CELL AND TISSUE ENGINEERING

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 研究中心，而研究中心不一定是研究者所在的机构。 该课程提供给参与合作项目的个人。培训的重点是所有基本技能的动手操作，使个人能够进行涉及使用培养细胞和/或工程组织的实验。 目前的培训包括：基本的无菌细胞培养技术、细胞保存和回收方法、细胞计数、Rose小室中的细胞培养、正常真皮和病理真皮来源的人体组织和人胶质母细胞瘤的解离以建立细胞系供将来使用、胶原蛋白和纤维蛋白中的肿瘤球体和筏培养。此外，我们还提供无壳鸡胚CAM（绒毛尿囊膜）模型与肿瘤球体植入的培训。TUNEL测定、肌动蛋白可视化和各种其他标记方法，其允许特定亚细胞器和中间丝的单独MPM或MPM/OCT成像，目的是监测治疗处理的结果和细胞外基质中细胞功能的动态。 在过去的一年里，我们发展了我们的培训课程。我们在2006年12月中旬收到了第一例人类胶质母细胞瘤活检，开始了肿瘤干细胞分离，特别关注人类胶质母细胞瘤CD 133阳性细胞（工作正在进行中）。获取建立无壳鸡胚CAM模型的过程的数字图像并用于训练。从其他机构接受培训的各方广泛要求提供这些图像。我们完整的筏式培养方案也被广泛要求。我们收到了许多请求，并提供了建立肿瘤球体的培训。在未来的一年里，我们将集中精力在网上发布所有现有的协议，并附上数字图像，供非现场合作者通过密码访问和内部自我指导培训。

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.