组织单腺体内单细胞的基因变异分析方法

发布时间：2019-01-28 09:42

【摘要】：从单细胞尺度进行细胞异质性的分析是深度理解细胞群体关系的关键。组织中的单细胞由于细胞类型不同,尺寸往往相差很大,但是目前常用的基于微孔板和Fluidigm公司的微流控的单细胞组学研究方法,需要入口的单细胞大小相近。本研究以胃组织为例,建立了一种组织单细胞的基因变异分析方法,实现了尺寸差异较大的单细胞的基因变异分析。在该方法中,先将胃组织裂解获得单个腺体,再将单个腺体酶解得到不同大小的腺体内单细胞,然后把这些单细胞铺在聚乙烯萘膜载玻片上,进行激光显微切割分选、全基因组放大,最后测其微卫星的长度。利用该方法,成功在肠上皮化生腺体内部检测到微卫星长度的变化,并灵活地对尺寸差异大的组织细胞以及肠化生腺体细胞进行了精细分析。此外,这种单细胞分析方法还可以对带有不同标记的细胞进行低通量和高通量的基因组分析,为单细胞尺度上的组织异质性研究提供了一种高度灵活的分析方法。
[Abstract]:The analysis of cell heterogeneity on a single cell scale is the key to a deep understanding of cell population relationship. The size of single cells in tissue varies greatly due to different cell types. However, the current microfluidic single cell research methods based on micropore plate and Fluidigm company require the same size of the single cell entrance. In this study, a method for the analysis of single cell gene variation in gastric tissue was established, and the genetic variation analysis of single cell with great difference in size was realized. In this method, a single gland is obtained by splitting the stomach tissue, and then the single cells of different sizes are obtained by enzymatic hydrolysis. Then, the single cells are laid on the polyethylene naphthalene film slide glass for laser microdissection and sorting. The whole genome was amplified and the length of the microsatellite was finally measured. Using this method, microsatellite length was detected successfully in intestinal metaplasia glands, and the fine analysis of large size difference tissue cells and intestinal metaplasia gland cells was carried out. In addition, this single cell analysis method can also carry out low throughput and high throughput genomic analysis of cells with different markers, which provides a highly flexible analysis method for the study of tissue heterogeneity on a single cell scale.
【作者单位】：上海交通大学系统生物医学研究院;上海交通大学医学院附属仁济医院;上海交通大学生物医学工程学院;
【基金】：国家自然科学重大研究计划项目(编号:91529302)资助~~
【分类号】：Q23

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