LASER CELL PROCESSING FOR BASIC AND CLINICAL RESEARCH

Photosis是一个融合了高速 生物样品的光学扫描、图像分析和计算机控制 激光照射样品内的特定目标，目的是 引发生物反应。要在混合的 种群由诸如大小、形状、荧光或 其他显著特征。一旦确定，单个细胞就成为目标 用激光来诱导所需的反应，如细胞死亡、光学修复 (用于基因转移)，甚至使特定的mRNA转录本失活 牢房。目前的Beta1-Prototyle系统可以处理数亿 在无菌条件下一小时内获得细胞，这使得它对几个 研究和临床应用。事实上，这个原型有几个 与其他细胞处理方法相比，如流式细胞术，以及 报告中显示的初步数据支持了这一结论。Photosis有 许多潜在的用途，这项提议汇集了许多机构 和研究人员调查并定义这一技术的可能应用 新技术。在其当前配置中，该仪器使用单个 用于细胞检测的颜色和用于在每个目标细胞中诱导坏死的激光。 这些能力使第一个应用成为可能，即清除肿瘤细胞 来自自体非霍奇金淋巴瘤干细胞移植，因为这种污染肿瘤 众所周知，细胞是导致疾病复发的因素。其他应用程序将 将被开发，其中一些将需要修改系统设计和 建造新的原型。原型将被放置在四个合作伙伴中 将进行基础和临床应用研究的地点， 包括：1)体内纯化干细胞亚群的研究 异种胎羊移植模型；2)评估非霍奇金淋巴瘤的人类临床试验 自体干细胞移植中的净化；3)遗传基因的纯化。 表达可选择转基因的修饰的干细胞和T细胞，以及 通过光学作用选择性地转导混合物中的特定细胞；以及 4)从纯化的原代人前列腺组织中获得准确的mRNA表达谱 癌细胞群体。 建议的商业应用： 这项拟议的工作将产生几种新型的生物工程仪器 以促进细胞处理的最先进水平。这些仪器将被使用 在这个推进干细胞生物学基础和临床研究的项目中， 免疫学和基因组学。一旦开发出来，由此产生的技术将是 在其他领域也很有用，其中一些在中进行了描述。

Photosis is a technology platform that incorporates high-speed optical scanning of biological samples, image analysis, and computer-controlled laser-irradiation of specific targets within the sample for the purpose of inducing a biological response. Specific cells to be treated within a mixed population are identified by parameters such as size, shape, fluorescence, or other distinguishing features. Once identified, individual cells are targeted with a laser to induce a desired response, such as cell death, optoporation (for gene transfer), or even inactivation of a specific mRNA transcript within the cell. The current beta1-prototyle system can process hundreds of millions of cells in an hour under sterile conditions, making it useful for several research and clinical applications. In fact, this prototype has several advantages over other methods of cell processing such as flow cytometry, and this conclusion is supported by preliminary data shown within. Photosis has many potential uses, and this proposal brings together a number of institutions and researchers to investigate and define the possible applications of this novel technology. In its current configuration, the instrument uses a single color for cell detection and a laser to induce necrosis in every targeted cell. These capabilities enable the first application, which is tumor cell purging from autologous NHL stem cell transplants, because such contaminating tumor cells are known to contribute to disease relapse. Additional applications will be developed, some of which will require modifications to the system design and building of new prototypes. The prototypes will be placed at four partnership sites where the basic and clinical applications research will be carried out, including: 1) in vivo study of purified stem cell subpopulations in the xenogeneic fetal sheep transplant model; 2) human clinical trials to assess NHL purging in autologous stem cell transplantation; 3) purification of genetically- modified stem cells and T-cells expressing a selectable transgene, as well as selective transduction of specific cells in a mixture via optoporation; and 4) accurate mRNA expression profiling from purified primary human prostate cancer cell populations. PROPOSED COMMERCIAL APPLICATION: The proposed work will result in several types of novel bioengineering instrumentation for advancing the state-of-the-art in cell processing. These instruments will be used in this program to advance basic and clinical research in stem cell biology, cancer, immunology, and genomics. Once developed, the resulting technology will be useful in other areas as well, some of which are described within.