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确定脱细胞化主动脉中的残留DNA - - 通过各种方法净化DNA的光基量测量

Stefan Pentzold¹, Mengjin Li¹, Ana Lucia Paz Hernandez¹

¹Department of General, Visceral and Vascular Surgery, Research Group Experimental Transplantation Surgery, Jena University Hospital, Friedrich Schiller University Jena, Drackendorfer Straße 1, D-07747, Jena, Germany.

Molecular biology reports

|July 8, 2025

概括

在脱细胞化组织中精确量化残留的双链DNA (dsDNA) 对患者的安全至关重要. 脱盐DNA后进行PicoGreen或QPCR,可以可靠地量化脱细胞化血管中的dsDNA.

科学领域:

生物材料科学生物材料科学
组织工程是组织工程.
分子生物学分子生物学

背景情况:

动物组织的脱细胞化产生了组织工程的支架.
脱细胞化组织中残留的双链DNA (dsDNA) 可以引起患者的不良反应.
精确的dsDNA量化是必不可少的,但由于净化方法的干扰,这是一项挑战.

研究的目的:

确定净化和量化脱细胞容器中的残留dDNA的最佳方法.
为了比较不同的DNA净化技术和量化测试.
为了确保临床应用的脱细胞化组织支架的安全性.

主要方法:

用具有或没有DNase I.的清洁剂去细胞化大鼠胸前动脉.
采用了三种DNA净化方法:组织溶解,盐分和基于二氧化的固体相.
使用PicoGreen,实时QPCR,凝电泳,紫外线光谱测量和DAPI染色来量化dsDNA.

主要成果:

DNase I 治疗显著降低了 dsDNA 的水平.
皮科格林和DAPI染色显示,脱细胞化后的dsDNA显著减少.
脱盐DNA提供了可靠的结果,使用PicoGreen,QPCR和凝电泳.
在这种情况下,发现光谱光度计对于dSDNA量化是不可靠的.

结论:

基于光的量化 (PicoGreen) 是多功能跨净化方法.
脱盐DNA是一种可靠的净化方法,用于随后的PicoGreen,QPCR和凝电泳.
不建议在脱细胞化组织中进行dsDNA量化时使用光谱光度计.
组织部分的DAPI染色提供了补充DNA量化.

关键词:

净化 DNA 的 DNA 净化.DNA定量化 DNA定量化 DNase I 是一个基因组.脱细胞化脱细胞化洗剂是一种洗剂.

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