PROLIFERATION AND DIFFERENTIATION OF ISOLATED STEM CELLS

分离的干细胞的增殖和分化

• 批准号: 3365640
• 负责人: JOHN W ADAMSON
• 金额: $ 15.95万
• 依托单位: NEW YORK BLOOD CENTER
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 1995-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3365640
• 关键词: cell differentiation; cell population study; cell sorting; colony stimulating factor; erythropoietin; extracellular matrix proteins; genetic transcription; growth factor receptors; hematopoietic growth factor; hematopoietic stem cells; human tissue; interleukin 1; interleukin 3; interleukin 6; laboratory mouse; laboratory rat; polymerase chain reaction; tissue /cell culture

The proposed studies will examine the effects of two novel hematopoietic growth factors - stem cell factor (SCF) and Rauscher factor - on the self- renewal and differentiation of highly enriched populations of murine and human hematopoietic stem cells (HSC). Recombinant purified rat SCF will be studied for its effect alone and with other growth factors such as IL-1, IL-3, IL-6, GM-CSF and erythropoietin (Epo) on the self renewal and differentiation of HSC isolated by physical means. Suspensions of HSC will be used in "read out" assays for the generation of colony-forming cells (CFC). In vivo, murine HSC function will be assayed by day 12 spleen colony-formation and long-term marrow reconstitution. The effects of recombinant human HSC will be studied using highly enriched populations of human progenitor cells (from marrow and umbilical cord blood) separated by flow cytometry or derived from blast cell colonies. SCF will be tested alone and in combination with other hematopoietic growth factors for generation of CFU-Mix and unilineage CFC in read-out assays. Rauscher factor, a potentially novel growth factor which is not species- specific, is most easily detectable when hematopoietic CFC come into contact with irradiated Rauscher cells. Thus, Rauscher cells may form a matrix containing Rauscher factor which can be studied by adding enriched murine or human SCF in low numbers to an established feeder. All cultures will be conducted under serum-deprived conditions to eliminate factors or co-factors provided in fetal bovine serum. These studies should provide insight into the mechanisms of action of these factors and the range of progenitor cells acted upon. The results also may allow a basis for the use of combinations of these and other factors to amplify HSC numbers appropriate for transplantation or quantities of differentiated cells appropriate for transfusion.

拟议的研究将检查两种新的造血作用， 生长因子-干细胞因子（SCF）和Rauscher因子-对自身的影响， 更新和分化高度富集的鼠和 人造血干细胞（HSC）。 重组纯化的大鼠SCF将 研究其单独作用和与其它生长因子如IL-1， IL-3、IL-6、GM-CSF和促红细胞生成素（Epo）对自我更新和 通过物理方法分离的HSC的分化。 HSC的暂停将 用于产生集落形成细胞的“读出”测定 （CFC）。 在体内，将通过第12天的脾脏测定鼠HSC功能 集落形成和长期骨髓重建。 重组人HSC的作用将使用高度富集的 人祖细胞群（来自骨髓和脐带 血液），或来源于胚细胞集落。 SCF将单独测试，并与其他造血生长因子联合测试。 用于在读出测定中产生CFU-Mix和单系CFC的因子。 劳舍尔因子，一种潜在的新型生长因子， 特异性，最容易检测时，造血CFC进入 与辐射过的Rauscher细胞接触。 因此，Rauscher细胞可以形成 含有Rauscher因子的矩阵，可以通过添加富集的 小鼠或人SCF以低数量添加到已建立的饲养者中。 所有文化 将在无血清条件下进行，以消除因素或 辅因子在胎牛血清中提供。 这些研究应提供深入了解这些药物的作用机制， 因子和祖细胞的作用范围。 结果也可能 允许使用这些因素和其他因素的组合的基础， 扩增适于移植的HSC数量或扩增适于移植的HSC数量。 适合输血的分化细胞。