Kolosov E V, Osipova L A, Kaz'min S D, Bykorez A I
Tsitologiia. 1984 Aug;26(8):968-73.
In different periods after partial hepatectomy (0, 11, 18, 24, 48 and 72 hours) isolated liver cell nuclei were separated according to their sizes using free sedimentation at 1 g in the exponential 0.1-1.0 M concentration gradient of sucrose at 4 degrees C. A sedimentogram of the intact liver cell nuclei displays three peaks corresponding to the nuclei of stromal cells, and diploid and tetraploid nuclei of hepatocytes. At 18 and 24 hours after the operation, but not at 0 or 11 hours, a curve of 3H-thymidine incorporation is represented by three maxima suggesting the influx of hepatocytes of different ploidy in the S-period. Then, 48 and 72 hours after the partial hepatectomy two additional peaks appear corresponding to 8- and 16-ploid nuclei, and a decrease in the portion of diploid nuclei is seen up to the full absence of the corresponding peak. Thus, the method of sedimentation at unit gravity of isolated nuclei using radioisotope labeling allows to obtain the information on the structure of the heterogeneous proliferating cell population.
在部分肝切除术后的不同时间段(0、11、18、24、48和72小时),通过在4℃下于0.1 - 1.0M蔗糖指数浓度梯度中以1g自由沉降,根据大小分离出分离的肝细胞核。完整肝细胞核的沉降图显示出三个峰,分别对应于基质细胞核以及肝细胞的二倍体和四倍体细胞核。在术后18和24小时,但不是在0或11小时,3H - 胸腺嘧啶核苷掺入曲线由三个最大值表示,表明不同倍性的肝细胞在S期流入。然后,在部分肝切除术后48和72小时出现另外两个对应于8倍体和16倍体细胞核的峰,并且二倍体细胞核部分减少直至相应峰完全消失。因此,使用放射性同位素标记在单位重力下对分离细胞核进行沉降的方法能够获得关于异质增殖细胞群体结构的信息。