Moir R D, Smith G J, Stewart B W
Cell Biol Int Rep. 1983 Mar;7(3):227-36. doi: 10.1016/0309-1651(83)90230-8.
Structural analysis of DNA from primary hepatocytes has been made by chromatography on benzoylated-DEAE-cellulose. DNA was labelled in vivo by incorporation of 3H-TdR after partial hepatectomy 2 weeks prior to animals being used to establish hepatocyte cultures. Structural change in DNA associated with the transition to primary culture was assessed by comparison of data with that obtained when DNA was isolated from the intact organ immediately after death. In terms of this comparison, DNA from primary hepatocytes was bound more extensively to benzoylated-DEAE-cellulose in the presence of 1.0 M NaCl. Thus 32% of primary hepatocyte DNA was recovered in the formamide-eluted fraction after chromatography, a result which regions. This proportion of formamide-eluted DNA in primary hepatocytes could not be further reduced by extensive shearing and could not be affected by manipulation of the isolation procedure. Analysis of DNA isolated from hepatocyte cell lines revealed structural characteristics similar to DNA isolated from intact liver. The data imply that extensive structural damage to DNA is a consequence of the primary status of the hepatocytes examined.
通过在苯甲酰化二乙氨基乙基纤维素上进行色谱分析，对原代肝细胞的DNA进行了结构分析。在用于建立肝细胞培养的动物前2周部分肝切除后，通过掺入3H-TdR在体内对DNA进行标记。通过将数据与动物死后立即从完整器官中分离DNA时获得的数据进行比较，评估了与向原代培养转变相关的DNA结构变化。根据这种比较，在1.0M NaCl存在下，原代肝细胞的DNA与苯甲酰化二乙氨基乙基纤维素的结合更为广泛。因此，色谱分析后，32%的原代肝细胞DNA以甲酰胺洗脱级分的形式回收，这一结果……。原代肝细胞中甲酰胺洗脱DNA的这一比例不会因广泛剪切而进一步降低，也不会受到分离程序操作的影响。对从肝细胞系中分离的DNA的分析揭示了与从完整肝脏中分离的DNA相似的结构特征。数据表明，DNA的广泛结构损伤是所检查肝细胞原代状态的结果。
