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在组织培养基中,利用等动力学密度梯度的菲可分离具有组织化学可证实的葡萄糖-6-磷酸酶的仓鼠肾细胞悬浮液中的细胞。

Separation of cells having histochemically demonstrable glucose-6-phosphatase from suspensions of hamster kidney cells in an isokinetic density gradient of Ficoll in tissue culture medium.

作者信息

Pretlow T G, Jones J, Dow S

出版信息

Am J Pathol. 1974 Feb;74(2):275-86.

Abstract

Suspensions of enzymatically disaggregated hamster kidney cells were separated primarily according to differences in diameter using velocity sedimentation in a previously described isokinetic density gradient and according to differences in density in an isopycnic density gradient. Cells which contained histochemically demonstrable glucose-6-phosphatase were thought to be cells from proximal tubules and constituted 46.5 +/- 14.1% of the cells in the starting sample suspension of disaggregated kidney cells. The purest gradient fractions from experiments using velocity sedimentation contained 98.0 +/- 0.6% cells which demonstrated glucose-6-phosphatase activity. More than 99.0% of these cells excluded trypan blue. Isopycnic sedimentation was not an effective means of purifying proximal tubule cells.

摘要

使用先前描述的等速密度梯度中的速度沉降法,根据直径差异对酶解的仓鼠肾细胞悬液进行初步分离;并使用等密度密度梯度,根据密度差异进行分离。含有组织化学可证实的葡萄糖-6-磷酸酶的细胞被认为是来自近端小管的细胞,在起始的肾细胞解离样品悬液中占细胞总数的46.5±14.1%。使用速度沉降法进行实验得到的最纯梯度级分中,有98.0±0.6%的细胞表现出葡萄糖-6-磷酸酶活性。这些细胞中有超过99.0%的细胞排斥台盼蓝。等密度沉降不是纯化近端小管细胞的有效方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3dbd/1910781/37b2547bf450/amjpathol00478-0081-a.jpg

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