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（改编自申请人摘要） 转基因和基因敲除小鼠的表型筛选，以确定是否存在以下疾病： 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，由于小鼠的造血系统相对容易， 分离、操作和表征，它提供了有价值模型系统， 研究转基因和基因敲除在一般生物学中的作用 流程. 这些研究人员最近开发了一种半自动的高 基于流式细胞术的通量筛选技术来评估效果 人造血干细胞自身的替代培养条件 更新和分化。 这项技术，称为流式细胞术高 发射量筛选（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。 他们建议修改这一点 技术，以便将其用作高通量筛选技术， 表征转基因和敲除小鼠的造血特征。 为了 为了使F-HTS适合作为鼠系统中的筛选工具，他们提出： (1)通过仪器和软件使F-HTS技术更加自动化 （2）建立F-HTS染色法定量检测小鼠骨髓， 淋巴细胞和干细胞，以及用于测量细胞内细胞因子 （3）验证F-HTS技术在代表性的 一组野生型和转基因小鼠，以及（4）开发网页， 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技术用于高效筛选大量诱变小鼠， 造血系统的异常，并可适用于 也会筛选其他组织 (End抽象）。