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从正常大鼠和经致癌物处理的大鼠中分离肝细胞的分级分离。

The fractionation of isolated liver cells from normal and carcinogen treated rats.

作者信息

Horsfall A C, Ketterer B

出版信息

Br J Cancer. 1976 Jan;33(1):96-104. doi: 10.1038/bjc.1976.10.

Abstract

Suspensions of isolated cells were obtained from livers of normal rats and rats treated with the hepatocarcinogen N,N-dimethyl-4-aminoazobenzene. Differential centrifugation of dispersed cells yielded a large parenchymal cell fraction and a small non-parencymal cell fraction. By means of rate sedimentation through different concnetrations of Ficoll, parenchymal cells were separated into cells with fast, intermediate and slow rates of sedimentation. Periods of sedimentation were brief and centrifugal forces low in order to retain the best possible state of preservation of cells. DNA, RNA and protein contents, acid phosphatase activity, cell size and nucleocytoplasmic ratios of parenchymal cells sedimenting at fast, intermediate and slow rates were measured. Cell fractions from normal livers had properties suggesting that faster sedimenting cells were derived from the centre and middle of the lobule whereas slowly sedimenting cells were periportal; however, much of the periportal cell population remained in a residue of undissociated tissue. Compared with normal cells, carcinogen treated cells appeared to fractionate according to different physical and chemical criteria and could not be related to their origin within the liver lobule. They were smaller, slower sedimenting, lower in protein and RNA content and acid phosphatase activity. The tissue residue contained abnromal histological structures.

摘要

从正常大鼠以及用肝癌诱发剂N,N - 二甲基 - 4 - 氨基偶氮苯处理过的大鼠的肝脏中获取分离细胞悬液。对分散的细胞进行差速离心,得到一个大的实质细胞部分和一个小的非实质细胞部分。通过在不同浓度的菲可中进行速率沉降,实质细胞被分离成沉降速率快、中等和慢的细胞。沉降时间短暂,离心力较小,以便尽可能保持细胞的最佳保存状态。测量了以快、中、慢速率沉降的实质细胞的DNA、RNA和蛋白质含量、酸性磷酸酶活性、细胞大小和核质比。正常肝脏的细胞部分具有的特性表明,沉降较快的细胞来自小叶的中央和中间部分,而沉降较慢的细胞是门静脉周围的;然而,许多门静脉周围的细胞群体仍留在未解离组织的残余物中。与正常细胞相比,经致癌物处理的细胞似乎根据不同的物理和化学标准进行分级,并且与它们在肝小叶内的起源无关。它们更小,沉降更慢,蛋白质和RNA含量以及酸性磷酸酶活性更低。组织残余物含有异常的组织结构。

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