Generation of Hematopoietic Stem Cells from Induced Pluripotent Stem Cells

从诱导多能干细胞产生造血干细胞

• 批准号: 8710195
• 负责人: MARCUS O MUENCH
• 金额: $ 26.36万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8710195
• 关键词: Address; Adult; Autologous Stem Cell Transplantation; Blood; Cell Line; Cells; Chromosome abnormality; Commit; Development; Elements; Fetal Tissues; Fostering; Gene Expression; Gene Expression Profile; Generations; Genes; Globin; Goals; Growth; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hemoglobinopathies; Human; Human Engineering; Immunodeficient Mouse; In Vitro; Individual; Instruction; Maintenance; Mesoderm; Methods; Mus; Outcome; Patients; Play; Pluripotent Stem Cells; Procedures; Process; Production; Reproducibility; Role; Safety; Staging; Stem Cell Development; Stromal Cells; Technology; Teratoma; Testing; Therapeutic; Umbilical Cord Blood; base; cell growth; cytokine; embryonic stem cell; fetal; hematopoietic tissue; induced pluripotent stem cell; leukemia; peripheral blood; progenitor; programs; reconstitution; research study; response; stem; technology development

PROJECT SUMMARY (See instructions): The overall goal of Project 3 is to develop a method to efficiently and reproducibly differentiate induced pluripotent stem (IPS) cells into hematopoietic stem cells (HSCs). Aim 1 of this project will determine the best conditions for the mesodermal differentiation of iPS cells and the generation of HSCs. This aim will test the hypothesis that an engineered human stromal cell line offers the best method to reliably differentiate IPS cells into hematopoietic precursors. Experiments will compare methods of differentiating IPS cells using embryoid bodies (EB) cultures and stromal cell lines to support the differentiation of IPS cells into HSCs. Various elements of these two differentiation methods will be studied to determine the best method to promote mesodermal differentiation and HSC creation. The second aim is to determine the optimal cytokine conditions for the survival, growth and expansion of HSCs generated from iPS cells. This aim will test the hypothesis that HSCs generated from IPS are more similar in their cytokine responses to fetal HSCs than adult HSCs. Accumulation of HSCs in culture requires conditions that favor their survival and minimize their differentiation into committed progenitors. Various cytokines known to play a role in the early stages of hematopoiesis will be tested in combination to compare the similarity of HSCs generated from iPS cells to those isolated from fetal tissues, umbilical cord blood and adult peripheral blood. These experiments will optimize culture conditions for the production of HSCs. The third aim will determine the variability in the capacity of different IPS cell lines to differentiate into HSCs. This aim will test the hypothesis that different iPS cells lines are similarly capable to form HSCs that can generate long-term multilineage reconstitution in immunodeficient mice. This aim will test isolated HSCs derived from multiple IPS cell lines for their capacity to provide long-term reconstitution without teratoma or leukemia formation, chromosomal abnormalities or other obvious functional deficiencies. The outcome of the experiments will be development of technology to efficiently generate transplantable HSCs from genetically corrected IPS cells.

项目摘要（参见说明）： 项目3的总体目标是开发一种有效且可重复地将诱导多能干细胞（IPS）分化为造血干细胞（HSC）的方法。该项目的目标 1 将确定 iPS 细胞中胚层分化和 HSC 生成的最佳条件。这一目标将检验以下假设：工程化人类基质细胞系提供了可靠地将 IPS 细胞分化为造血前体细胞的最佳方法。实验将比较使用胚状体 (EB) 培养物和基质细胞系分化 IPS 细胞的方法，以支持 IPS 细胞分化为 HSC。 将研究这两种分化方法的各个要素，以确定促进中胚层分化和 HSC 生成的最佳方法。第二个目标是确定 iPS 细胞生成的 HSC 存活、生长和扩增的最佳细胞因子条件。这一目标将检验以下假设：IPS 产生的 HSC 与胎儿 HSC 的细胞因子反应比成人 HSC 更相似。 HSC 在培养物中的积累需要有利于其生存并尽量减少其分化为定向祖细胞的条件。将组合测试已知在造血早期发挥作用的各种细胞因子，以比较 iPS 细胞产生的 HSC 与从胎儿组织、脐带血和成人外周血中分离的 HSC 的相似性。这些实验将优化 HSC 生产的培养条件。第三个目标将确定不同 IPS 细胞系分化为 HSC 的能力的差异。这一目标将检验以下假设：不同的 iPS 细胞系同样能够形成 HSC，从而在免疫缺陷小鼠中产生长期的多谱系重建。这一目标将测试从多种 IPS 细胞系分离的 HSC 的长期重建能力，而不会形成畸胎瘤或白血病、染色体异常或其他明显的功能缺陷。实验的结果将是开发从基因校正的 IPS 细胞中有效生成可移植 HSC 的技术。