PROLIFERATION AND DIFFERENTIATION OF ISOLATED STEM CELLS

拟议中的研究将检验两种新的造血剂的效果 生长因子-干细胞因子(SCF)和劳谢尔因子-对自身的影响 高度丰富的小鼠和小鼠群体的更新和分化 人造血干细胞(HSC)。重组纯化的大鼠干细胞将被 研究了它的单独作用以及与其他生长因子如IL-1的作用， IL-3、IL-6、GM-CSF和EPO对自我更新和 物理方法分离的HSC的分化。暂停HSC将会 用于集落形成细胞的“读出”检测 (Cfc)。在体内，小鼠的HSC功能将在第12天进行脾功能检测 集落形成和长期的骨髓重建。 重组人HSC的作用将用高度浓缩的 人的祖细胞(来自骨髓和脐带)的群体 血液)通过流式细胞术分离的或来自原始细胞集落的。 干细胞干细胞将单独进行测试，并与其他造血生长一起进行测试。 读出化验中产生CFU混合和单线CFC值的因素。 Rauscher因子，一种潜在的新的生长因子，它不是物种- 特异性，是最容易检测到的当造血型氟氯化碳进入 接触受辐射的劳舍尔细胞。因此，Rauscher细胞可以形成一种 含有Rauscher因子的矩阵可以通过添加浓缩来研究 将数量较少的小鼠或人类干细胞移植到已建立的饲养层。所有文化 将在缺乏血清的条件下进行，以消除因素或 胎牛血清中提供的辅助因子。 这些研究应该为深入了解这些药物的作用机制提供帮助 因素和祖细胞作用的范围。结果还可能 为使用这些因素和其他因素的组合提供基础 放大适合移植的HSC数量或数量 适合输血的分化细胞。

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.