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用于中期染色体分析和染色体微阵列分析的绒毛膜绒毛样本制备。

Preparation of chorionic villus samples for metaphase chromosome analysis and chromosomal microarray analysis.

作者信息

Breman Amy, Patel Ankita

机构信息

Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas, USA.

出版信息

Curr Protoc Hum Genet. 2012 Oct;Chapter 8:Unit8.3. doi: 10.1002/0471142905.hg0803s75.

Abstract

Chorionic villi are composed of an outer layer of trophoblastic cells and an inner mesenchymal cell core. They can be prepared for chromosome analysis using a culture method wherein villi are disaggregated by mechanical and enzymatic methods and the resulting cell suspension is used to establish primary cultures. Mesenchymal cells of the villus core are released by this procedure and the fibroblasts are actively proliferative in tissue culture. Cultures can be used for cytogenetic analysis after ∼1 week. Slides prepared by this technique can be stained using trypsin-Giemsa banding and analyzed for chromosomal abnormalities in fetal tissue. Chorionic villi may also be assessed by chromosomal microarray analysis (CMA). For this purpose, a method for extraction of high-quality DNA from CVS tissue is also described here.

摘要

绒毛膜绒毛由外层滋养层细胞和内层间充质细胞核心组成。可以使用一种培养方法来制备用于染色体分析的样本,该方法通过机械和酶促方法将绒毛解离,所得细胞悬液用于建立原代培养。通过该程序可释放绒毛核心的间充质细胞,并且成纤维细胞在组织培养中具有活跃的增殖能力。培养物在约1周后可用于细胞遗传学分析。用该技术制备的玻片可使用胰蛋白酶-吉姆萨显带染色,并分析胎儿组织中的染色体异常。绒毛膜绒毛也可通过染色体微阵列分析(CMA)进行评估。为此,本文还描述了一种从绒毛取样组织中提取高质量DNA的方法。

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