DNA Methylation Analysis as a Tool for Cell Typing
DNA Methylation Analysis as a Tool for Cell Typing
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DOI：
10.4161/epi.1.1.2643
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发表时间：
2006-01-01
期刊：
影响因子：
3.7
通讯作者：
Olek, Sven
中科院分区：
文献类型：
--
作者：
Baron, Udo;Tuerbachova, Ivana;Olek, Sven
细胞治疗方法目前缺乏明确的质量控制措施，以保证临床应用的安全性，并为监管部门的批准创造一致的标准。这些方法依赖于供体细胞的分离、纯化和可能的离体操作。由于这些细胞暴露在人工环境中，有可能偏离自然生长过程。细胞培养的异质性是一个固有的问题。因此，验证细胞身份和定量亚群是强制性的。专注于培养的人类原代细胞，我们测试了DNA甲基化模式是否作为独特的细胞类型标记。我们确定了细胞类型特异性差异甲基化基因区域（CDMs）的面板，这些区域为这些细胞类型产生明确的轮廓。应用甲基化敏感的单核苷酸引物扩展产生二元细胞类型描述符（“条形码”），允许定量细胞混合物。因此，基于甲基化的分析表明，它们是体外操作细胞的表征和质量控制的有前途的工具。
Cell therapeutic approaches currently lack definitive quality control measures which guarantee safety in clinical applications and create consistent standards for regulatory approval. These approaches rely on isolation, purification and possibly ex vivo manipulation of donor cells. Since such cells are exposed to artificial environments, there is potential for deviations from natural growth processes. The resulting heterogeneity of cell cultures is an inherent problem. Therefore, verification of cell identity and quantification of subpopulations is mandatory. Focusing on cultured human primary cells, we tested whether DNA methylation patterns serve as distinctive cell type markers. We identified panels of cell type specific differentially methylated gene regions (CDMs) which produce unambiguous profiles for these cell types. Applying methylation sensitive single nucleotide primer extension generated binary cell type descriptors ("barcodes") which allow quantification of cell mixtures. Thus, methylation based analytics suggest themselves as promising tools for the characterization and quality control of ex vivo manipulated cells.