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（改编自申请人的摘要） 转基因和基因敲除小鼠的表型筛选 造血细胞的自我更新、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。 另外，由于小鼠的造血系统相对容易 隔离、操纵和表征，它提供了一个有价值的模型系统 研究转基因和基因敲除对一般生物制剂的作用 流程。 这些研究人员最近开发了一种半自动化的高 基于流式细胞术的通量筛选技术评估效果 替代培养条件对人类造血干细胞自身的影响 更新和差异化。 该技术称为流式细胞术高 通量筛选（F-HTS）可以高效、准确地表征 造血生物学的关键方面，包括细胞增殖， 分化、自我更新和细胞凋亡。 他们建议修改这个 技术，以便将其用作高通量筛选技术 描述转基因和基因敲除小鼠的造血特征。 为了 为了将 F-HTS 用作小鼠系统中的筛选工具，他们建议： (1) 通过仪器和软件使F-HTS技术更加完全自动化 另外，(2) 开发用于定量小鼠骨髓的 F-HTS 染色方法， 淋巴细胞和干细胞，以及用于测量细胞内细胞因子 生产和细胞凋亡，（3）在代表性中验证 F-HTS 技术 野生型和转基因小鼠小组，以及（4）开发一个网页 传播有关 F-HTS 技术和结果的信息 正在进行的研究。 这些项目将为使用F- HTS 技术可有效筛选大量诱变小鼠 造血系统异常，可适用于 也筛查其他组织。 （摘要结束。）