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首页> 外文学位 >Human adult stem cells (hMSC) from bone marrow stroma: Characterization and standardization of culture preparation for in vivo applications, and a possible new application ofhMSCs as a model system for differentiation.
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Human adult stem cells (hMSC) from bone marrow stroma: Characterization and standardization of culture preparation for in vivo applications, and a possible new application ofhMSCs as a model system for differentiation.

机译：来自骨髓基质的人类成体干细胞（hMSC）：用于体内应用的培养制备的表征和标准化，以及hMSC作为分化模型系统的可能的新应用。

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摘要

For in vivo and clinical applications of human adult stem cells from bone marrow stroma (hMSCs), culture preparation standards are needed to ensure that preparations of cells are suitable for use. During previous work carried out to characterize hMSCs, a colony-forming unit (CFU) assay was used to demonstrate that the clonal efficiency of culture preparations correlated with their multi-potentiality. Additionally, CFU assays were useful for optimizing the culture conditions for expanding hMSCs because the conditions that preserved the %CFUs, could be identified and adopted relatively quickly. However, the CFU assay is inherently retrospective in scope. It requires 2 weeks to get the result and determine if a sample has a high enough CFU to meet quality standards. In clinical applications the outcome of quality assessment might need to be known for the very preparation that is to be used just before it is administered. For these reasons a flow cytometric measurement was developed that can predict which cell samples will have higher CFU efficiency and therefore greater multi-potentiality.;Additionally, an analysis of gene expression was carried out to identify genes that were co-expressed during the time course of differentiation of hMSCs into chondrocytes or adipocytes using three different differentiation protocols. Co-expressed genes were examined to identify new target genes for transcription factors that drive differentiation of hMSCs. Hierarchical clustering of differentially expressed genes was carried out to define groups of similarly behaving genes within each experiment. Next, groups of genes that were co-expressed in all three differentiation experiments were defined. The data were used to identify new targets of transcription factors involved in the differentiation of human cells into adipocytes and chondrocytes by introducing a proportionality criterion into our definition of co-regulation. Co-regulation was defined more stringently by requiring a highly significant linear regression between the levels of expression of known targets of transcription factors that drive chondrogenesis or adipogenesis and additional co-expressed genes not previously shown to be targets of the same transcription factors. Using this approach we identified 91 genes that are candidate downstream targets for SOX9, SOX5, C/BEP-α or PPAR-γ.
机译：对于来自骨髓基质（hMSC）的人类成体干细胞的体内和临床应用，需要培养准备标准以确保适合使用的细胞准备。在表征hMSC的先前工作中，菌落形成单位（CFU）分析用于证明培养物的克隆效率与其多潜能相关。另外，CFU测定法可用于优化扩增hMSC的培养条件，因为保存％CFU的条件可以相对较快地识别和采用。但是，CFU分析在范围上本质上是回顾性的。需要2个星期才能得到结果，并确定样品的CFU是否足够高以符合质量标准。在临床应用中，对于即将使用的准备工作，可能需要知道质量评估的结果。由于这些原因，开发了一种流式细胞仪测量方法，可以预测哪些细胞样品将具有更高的CFU效率并因此具有更大的多能性。;此外，对基因表达进行了分析，以鉴定在时间过程中共表达的基因三种不同分化方案将hMSCs分化为软骨细胞或脂肪细胞的研究。检查共表达的基因以鉴定驱动hMSC分化的转录因子的新靶基因。进行差异表达基因的分级聚类，以定义每个实验中行为相似的基因的组。接下来，定义了在所有三个分化实验中共表达的基因组。通过将比例标准引入我们的共调节定义中，这些数据被用于识别参与人类细胞分化为脂肪细胞和软骨细胞的转录因子的新靶标。通过要求驱动软骨形成或脂肪生成的转录因子的已知靶标的表达水平与以前未显示为相同转录因子的靶标的其他共表达基因之间的高度显着线性回归，可以更严格地定义共调节。使用这种方法，我们确定了91个基因，它们是SOX9，SOX5，C /BEP-α或PPAR-γ的候选下游靶标。

著录项

  • 作者

    Smith, Jason Robert.;

  • 作者单位

    Tulane University.;

  • 授予单位 Tulane University.;
  • 学科 Biology Molecular.;Biology Cell.
  • 学位 Ph.D.
  • 年度 2004
  • 页码 143 p.
  • 总页数 143
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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