EXPANSION OF HUMAN STEM CELLS WITH TRANSPLANTATION MODEL

通过移植模型扩增人类干细胞

• 批准号: 2683727
• 负责人: RONALD L BROWN
• 金额: $ 37.14万
• 依托单位: QUALITY BIOLOGICAL, INC.
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2683727
• 关键词: cell cycle; cell population study; colony stimulating factor; hematopoietic stem cells; hematopoietic tissue transplantation; human tissue; mass tissue /cell culture; method development; sheep

DESCRIPTION (Adapted from applicant's abstract): The overall goal of this project is to develop the reagents and culture conditions devoid of serum that will support the ex vivo expansion of hematopoietic stem cells (HSC) for short-term and long-term engraftment. The studies presented here will evaluate the culture conditions necessary to retain HSC capable of in vivo short-term and long-term repopulating activity using the recently development sheep in utero transplantation model for human hematopoiesis. Feasibility data presented in this proposal show that CD34+ cells from human bone marrow cultured in serum-free medium containing low levels of cytokines retained the long-term repopulating cells. In contrast, CD34+ cells cultured under similar conditions, but in the presence of serum, exhibited only short-term marrow repopulating activity. In the Phase I effort, the investigator proposes to extend these studies comparing the culture conditions devoid of serum with serum containing culture condition. The applicants propose to analyze the cell populations from each culture condition for 1) cell cycle status, 2) viable cell number, 3) GM-CFC, and 4) population dynamics of the more primitive HSC versus differentiation of the myeloid cells using immunophenotype analysis. Such information will be valuable in correlating the culture condition necessary to maintain and those that deplete the population of cells necessary for long-term engraftment. The Phase II proposal will be to expand these studies to include cord blood normal human bone marrow and mobilized peripheral blood stem cells and to develop and optimize the culture reagents and conditions devoid of serum, necessary for ex vivo expansion of the long- term engrafting cells. The Phase II effort will focus on cytokine mixtures, concentration effects and times of exposure to expand the long-term engrafting cells. Such studies will be available in utilizing ex vivo culture in clinical protocols for the treatment of disease, and will be invaluable in designing the clinical regimens for ex vivo culture of HSC for transplantation.

描述（改编自申请人的摘要）：总体目标 该项目旨在开发缺乏的试剂和培养条件 支持造血干体外扩张的血清 细胞（HSC）用于短期和长期植入。 研究 这里介绍的将评估保留所需的培养条件 HSC 具有体内短期和长期增殖活性 使用最近开发的羊子宫内移植模型 人类造血。 本提案中提供的可行性数据显示 人骨髓中的 CD34+ 细胞在无血清培养基中培养 含有低水平的细胞因子保留了长期的再增殖 细胞。 相比之下，CD34+细胞在相似的条件下培养，但是 在血清存在的情况下，仅表现出短期的骨髓再生 活动。 在第一阶段的工作中，研究人员建议延长 这些研究比较了无血清培养条件和 含有血清的培养条件。申请人建议分析 1) 细胞周期的每种培养条件下的细胞群 状态，2) 活细胞数量，3) GM-CFC，以及 4) 群体动态 更原始的 HSC 与骨髓细胞的分化 使用免疫表型分析。 此类信息将很有价值 将维持所需的培养条件与那些 耗尽长期植入所需的细胞群。 第二阶段提案将扩大这些研究范围，将绳索纳入其中 血液 正常人骨髓和动员的外周血干 细胞并开发和优化培养试剂和条件 不含血清，是长期离体扩增所必需的 移植细胞。 第二阶段的工作将集中于细胞因子混合物， 浓度效应和暴露时间以扩大长期 移植细胞。 此类研究可利用离体 培养在疾病治疗的临床方案中，并将 对于设计 HSC 离体培养的临床方案非常有价值 用于移植。