Better ways to make Induced Pluripotential Stem (iPS) Cells

制造诱导多能干 (iPS) 细胞的更好方法

基本信息

批准号：
7685933
负责人：
JOHN W DAY
金额：
$ 8.18万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7685933
关键词：
Cancer Etiology Cartilage Cell Differentiation process Cell Line Cell Therapy Cells Charge Chromatin Clinical Defect Development Disease Effectiveness Elements Familiarity Fibroblasts Flow Cytometry Future Gene Combinations Gene Delivery Generations Genes Genome Goals Hereditary Disease Human Human Genetics Individual Insertional Mutagenesis Institutes Intervention Laboratories Life Methods Minnesota Mus Muscle Muscular Dystrophies Mutation Myopathy Oncogenic Pathology Pathway interactions Patients Pharmaceutical Preparations Phenotype Positioning Attribute Preparation Primary Cell Cultures Production Research Risk Sickle Cell Anemia Skeletal Muscle Skeletal system Staging Stem cells Technology Tissues Universities Viral Vector Virus Work base bone cell type embryonic stem cell experience human embryonic stem cell improved induced pluripotent stem cell new technology novel strategies rapid technique repository skills stem success transcription factor

项目摘要

Induced pluripotent stem cells (IPS cells) promise to revolutionize the study of muscle disease. iPS cells, made by introducing transcription factors into primary cell cultures, resemble embryonic stem cells, and can be used to study developmental mechanisms, their alteration in disease states, and to investigate the effects of drugs to correct the defect. Our scientific goal in this proposal is to develop methods for rapid creation of non-genetically modified iPS cells from selected patients with identified muscle disease. Success with these methods will improve efficiency and effectiveness of generation of iPS cells from specific disease fibroblasts maintained in Clinical Repository Research Core. These will be used to study the developmental pathology of these muscle diseases and to lay the basis for future cell therapy applications. An element of phenotypic characterization of the cells during differentiation will use the Force Assessment Research Core to determine whether identified muscle disease mutations alter force generation compared to wild type cells treated similarly. Specific Aims of the proposal are: Specific aim 1: To prepare iPS cells using a non-integrating viral vector. The working hypothesis is that integration into the genome is not required to bring about reprogramming to the iPS phenotype. Specific aim 2: To substitute less hazardous components in the preparation of iPS cells. The working hypothesis is that chromatin opening, rather than oncogenic transformation, is required to enable reprogramming to the iPS phenotype.

诱导多能干细胞（IPS 细胞）有望彻底改变肌肉疾病的研究。 iPS细胞，通过将转录因子引入原代细胞培养物中而制成，类似于胚胎干细胞，并且可以用于研究发育机制、它们在疾病状态下的改变，并研究其影响药物来纠正缺陷。我们在本提案中的科学目标是开发快速创建来自选定患有肌肉疾病的患者的非转基因 iPS 细胞。有了这些就成功了方法将提高从特定疾病成纤维细胞生成 iPS 细胞的效率和有效性维护在临床存储库研究核心中。这些将用于研究发育病理学这些肌肉疾病的研究并为未来的细胞治疗应用奠定基础。表型的一个要素细胞在分化过程中的表征将使用力评估研究核心来确定与治疗的野生型细胞相比，确定的肌肉疾病突变是否会改变力量的产生相似地。该提案的具体目标是：具体目标 1：使用非整合病毒载体制备 iPS 细胞。工作假设是不需要整合到基因组中即可重编程为 iPS 表型。具体目标 2：在 iPS 细胞的制备过程中替代危险性较低的成分。工作中假设是需要染色质开放而不是致癌转化来实现重编程为 iPS 表型。