FACS High Throughout Screening of Hematopoiesis

FACS 高通量造血筛查

• 批准号: 6656246
• 负责人: Doug Cress
• 金额: $ 30.18万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6656246
• 关键词: Internet; biomedical automation; biomedical equipment development; biotechnology; cell differentiation; cell proliferation; computer program /software; flow cytometry; gene targeting; genetic screening; genetically modified animals; hematopoiesis; hematopoietic stem cells; high throughput technology; information dissemination; laboratory mouse; lymphoblast; myeloid stem cell; stainings

DESCRIPTION (Adapted from the applicant's abstract) Phenotypic screening of transgenic and knockout mice for disorders in hematopoietic cell self renewal, differentiation and function will provide useful insights into the genetic basis of hematopoietic biology and diseases. In addition, because the murine hematopoietic system is relatively easy to isolate, manipulate and characterize, it provides a valuable model system for investigating the role of transgenes and gene knockouts on general biologic processes. These investigators have recently developed a semi-automated high throughput screening technique based on flow cytometry to evaluate the effects of alternative culture conditions on human hematopoietic stem cell self renewal and differentiation. This technique, termed Flow Cytometric High Throughput Screening (F-HTS) can efficiently and accurately characterize critical aspects of hematopoietic biology including cell proliferation, differentiation, self renewal and apoptosis. They propose to modify this technique in order to use it as a high throughput screening technique to characterize hematopoietic features of transgenic and knockout mice. In order to adapt the F-HTS as a screening tool int he murine system, they propose to: (1) more fully automate the F-HTS technique through instrument and software aditions, (2) develop F-HTS staining methods for quantitating murine myeloid, lymphoid and stem cells, as well as for measuring intracellular cytokine production and apoptosis, (3) validate the F-HTS technique in a representative panel of wild type and transgenic mice, and (4) develop a web page to disseminate information regarding the F-HTS technique and the results of ongoing studies. These projects will provide the foundation for using the F- HTS technique for efficiency screening large numbers of mutagenized mice for abnormalities of the hematopoietic system and may be adaptable for use in screening other tissues as well. (End of Abstract.)

描述（根据申请人的摘要改编） 转基因和基因敲除小鼠的表型筛查中的疾病 造血细胞自我更新，分化和功能将提供 对造血生物学和疾病的遗传基础的有用见解。 此外，由于鼠造血系统相对容易 孤立，操纵和特征，它为 研究转基因和基因敲除对一般生物学的作用 过程。 这些研究人员最近开发了一个半自动 基于流式细胞仪的吞吐量筛选技术以评估效果 关于人造血干细胞自我的替代培养条件 更新和分化。 该技术称为流式细胞术高 吞吐量筛选（F-HTS）可以有效，准确地表征 造血生物学的关键方面，包括细胞增殖， 分化，自我更新和凋亡。 他们建议对此进行修改 为了将其用作高通量筛查技术 特征是转基因和基因敲除小鼠的造血特征。 为了 为了使F-HTs作为筛选工具在鼠系统中，他们建议： （1）通过仪器和软件更完全自动化F-HTS技术 ADITION，（2）开发F-HTS染色方法来定量鼠髓样， 淋巴样和干细胞，以及用于测量细胞内细胞因子 生产和凋亡，（3）验证代表性的F-HTS技术 野生类型和转基因小鼠的面板，（4）开发一个网页 传播有关F-HTS技术的信息和结果 正在进行的研究。 这些项目将为使用F-提供基础 HTS技术用于效率筛选大量诱变小鼠 造血系统异常，可以适应 还筛选其他组织。 （摘要结束。）