Slowly cycling cells and hair follicle stem cells

缓慢循环细胞和毛囊干细胞

• 批准号: 7664493
• 负责人: Tudorita Tumbar
• 金额: $ 37.71万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-16 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7664493
• 关键词: Biochemical Markers; Biological Assay; Biological Models; Burn injury; Cell Culture Techniques; Cell Cycle; Cell Cycle Kinetics; Cell Fraction; Cell Proliferation; Cell Separation; Cell Transplantation; Cell physiology; Cells; Characteristics; Chromosomes; DNA; DNA biosynthesis; Data; Development; Disease; Epithelial; Equilibrium; Future; Gene Expression; Genes; Genomics; Goals; Green Fluorescent Proteins; Growth; Hair; Hair follicle structure; Heterogeneity; Histone H2B; Homeostasis; In Vitro; Knowledge; Label; Lead; Light; Maintenance; Methods; Modeling; Molecular; Morphogenesis; Mus; Mutation; Natural regeneration; Normal tissue morphology; Pattern; Phenotype; Physiologic pulse; Population; Population Heterogeneity; Process; Property; Research Personnel; Skin; Sorting - Cell Movement; Stem cells; System; Testing; Tissues; Work; base; clinical application; daughter cell; in vivo; interest; loss of function; programs; reconstitution; regenerative; repaired; residence; segregation; self-renewal; skin regeneration; stem cell niche; tool

DESCRIPTION (provided by applicant): We are interested in elucidating the molecular mechanisms involved in tissue stem cell self-renewal and fate choice, in normal tissue development and homeostasis. Stem cells hold great promises for future therapies of numerous deadly diseases, yet their basic properties are not well understood. Progress in the field has been hampered by the difficulty of identifying tissue stem cells in the absence of specific biochemical markers. Increasing evidence suggested that stem cells are infrequently dividing cells, that could be identified in many mouse tissues as DMA label retaining cells (or LRCs). Recently we developed a strategy applicable to many self-regenerating tissues to pulse-label LRCs with the green fluorescent protein (GFP) fused to histone H2B. We have used the mouse skin and the hair follicle as model systems, since the relation between LRCs and skin epithelial stem cells had been investigated in most depth. In our preliminary work we tracked and isolated GFP LRCs and demonstrated that at least a fraction of these cells had a stem cell phenotype. In this proposal we describe in three Specific Aims strategies to examine in more detail the relation between LRCs and stem cells. We will: 1) classify LRCs into distinct sub- populations and investigate their sternness; 2) examine their special cell cycle properties; 3) characterize their distinct patterns of gene expression. To accomplish these goals we use in vivo cell tracking and in vitro cell sorting methods combined with a variety of assays which include: hair regeneration and skin wounding; cell culture and skin reconstitution; cell cycle kinetics; and microarrays. First, our data will have global relevance for the isolation and characterization of putative stem cells with LRCs properties from many tissues. Although we focus on the skin, our work will lead the way for similar approaches in many systems which lack the specific means to separate stem cells from other cells of their tissue of residence. Second, stem cell proliferation must be tightly controlled in tissues to regulate the balance between normal and abnormal growth. Our studies will unravel some of the mechanisms developed by putative SCs to control their proliferation, an important first step in understanding such perturbation in the diseased tissues. Finally, our work will provide some of the basic knowledge necessary for more efficient manipulation of skin stem cells for clinical applications such as cell transplantation on burn victims.

描述（由申请人提供）：我们有兴趣阐明在正常组织发育和稳态中涉及组织干细胞自我更新和命运选择的分子机制。干细胞对众多致命疾病的未来疗法具有巨大的承诺，但其基本特性尚不清楚。在没有特定的生化标记的情况下，难以鉴定组织干细胞的困难，该领域的进展受到了阻碍。越来越多的证据表明，干细胞很少分裂细胞，在许多小鼠组织中可以识别为DMA标记保留细胞（或LRC）。最近，我们制定了一种适用于许多自我再生组织的策略，用于脉冲标签LRC，其绿色荧光蛋白（GFP）融合到组蛋白H2B。我们已经使用了小鼠皮肤和毛囊作为模型系统，因为已经研究了LRCS和皮肤上皮干细胞之间的关系。在我们的初步工作中，我们跟踪和分离了GFP LRC，并证明了这些细胞中至少一部分具有干细胞表型。在此提案中，我们在三个特定的目标策略中描述了LRC和干细胞之间的关系。我们将：1）将LRC分类为不同的子种群，并研究其严厉； 2）检查其特殊的细胞周期特性； 3）表征其基因表达的独特模式。为了实现这些目标，我们在体内细胞跟踪和体外细胞分选方法中使用多种测定法，其中包括：头发再生和皮肤受伤；细胞培养和皮肤重建；细胞周期动力学；和微阵列。 首先，我们的数据将具有从许多组织中具有LRCS特性的假定干细胞分离和表征的全球相关性。尽管我们专注于皮肤，但我们的工作将导致许多系统中类似方法的方向，这些系统缺乏将干细胞与居住组织其他细胞分开的特定方法。其次，必须在组织中严格控制干细胞增殖，以调节正常生长和异常生长之间的平衡。我们的研究将揭示假定SC控制其增殖的某些机制，这是理解患病组织中这种扰动的重要第一步。最后，我们的工作将提供一些基本知识，以更有效地操纵皮肤干细胞来进行临床应用，例如在烧伤受害者上移植细胞。