Gibb W, Lavoie J C
Can J Physiol Pharmacol. 1986 Feb;64(2):213-7. doi: 10.1139/y86-032.
Collagenase-dispersed cells from human chorion laeve were examined on Percoll gradients. The 3 beta-hydroxysteroid dehydrogenase (a trophoblast marker) and steroid sulfatase activities of the cells were measured and a system was developed to isolate enriched preparations of the trophoblast cells. No cells were found to sediment at Percoll concentrations greater than 50%, and using continuous gradients of Percoll there appeared to be cells with different 3 beta-hydroxysteroid dehydrogenase (3 beta HSD): steroid sulfatase ratios sedimenting in different regions of the gradient. Cells with a high ratio were found in the denser region of the gradient. Continuous gradients provided inadequate separations of distinct populations of cells, thus to obtain a more reproducible system to isolate cells, discontinuous gradients of Percoll were studied. A discontinuous gradient composed of 5, 20, 40, and 60% Percoll was developed and three bands of cells were found sedimenting at the 20, 40 and 60% interfaces, respectively. The number and appearance of cells at the 20 and 60% interfaces varied from tissue to tissue. In contrast, the cells sedimenting at the 40% interface were less variable, a substantial number was found to be present in every tissue studied, they were similar in appearance to the trophoblast cells and had high 3 beta HSD:sulfatase ratios.
对来自人平滑绒毛膜经胶原酶分散的细胞进行了Percoll梯度分析。测定了细胞的3β-羟基类固醇脱氢酶（一种滋养层标志物）和类固醇硫酸酯酶活性，并开发了一种系统来分离富集的滋养层细胞制剂。未发现细胞在Percoll浓度大于50%时沉降，并且使用连续的Percoll梯度时，似乎具有不同3β-羟基类固醇脱氢酶（3βHSD）：类固醇硫酸酯酶比值的细胞在梯度的不同区域沉降。高比值的细胞出现在梯度的较密区域。连续梯度对不同细胞群体的分离效果不佳，因此为了获得更可重复的细胞分离系统，研究了Percoll不连续梯度。开发了一种由5%、20%、40%和60%Percoll组成的不连续梯度，分别在20%、40%和60%的界面处发现有三带细胞沉降。在20%和60%界面处细胞的数量和外观因组织而异。相比之下，在40%界面处沉降的细胞变异性较小，在所研究的每个组织中都发现有大量此类细胞，它们在外观上与滋养层细胞相似，并且具有高3βHSD：硫酸酯酶比值。
