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在威斯康星大学溶液中低温保存长达3天后,用乙二胺四乙酸(EDTA)分离的大鼠肝实质细胞的活力、贴壁效率和外源性代谢酶活性得到良好维持。

Viability, attachment efficiency, and xenobiotic metabolizing enzyme activities are well maintained in EDTA isolated rat liver parenchymal cells after hypothermic preservation for up to 3 days in University of Wisconsin solution.

作者信息

Oesch F, Abdel-Latif H, Diener B

机构信息

Institute of Toxicology, University of Mainz, Germany.

出版信息

In Vitro Cell Dev Biol Anim. 1995 Sep;31(8):590-4. doi: 10.1007/BF02634311.

Abstract

Rat liver parenchymal cells were isolated by EDTA perfusion and were subsequently purified by Percoll centrifugation. The freshly isolated liver cells had a mean viability of 95% as judged by trypan blue exclusion. Isolated liver parenchymal cells were then stored at 0 degrees C for up to 1 wk in University of Wisconsin solution (UW). During this hypothermic preservation, the viability was only slightly reduced to 92% after 1 d and to 85% after 3 d at 0 degrees C. Thereafter, the viability decreased rapidly. After cold storage for up to 3 d, it was possible to use the parenchymal liver cells either in short-term suspension or in cell culture. The attachment efficiency in cell culture was the same for freshly isolated liver cells (84%) and after 2 d cold preservation (81%). The cytochrome P450 content and the enzyme activities of soluble epoxide hydrolase, UDP-glucuronosyl transferase, phenol sulfotransferase, and glutathione S-transferase were not significantly different between freshly isolated cells and cells after 3 d of hypothermic preservation. Furthermore, freshly isolated and intact liver cells stored for 3 d were used in the cell-mediated Salmonella mutagenicity test as a metabolizing system. Both fresh and stored liver parenchymal cells metabolized benzo(a)pyrene,2-aminoanthracene, and cyclophosphamide to their ultimate mutagens.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

通过乙二胺四乙酸(EDTA)灌注分离大鼠肝实质细胞,随后通过聚蔗糖(Percoll)离心进行纯化。通过台盼蓝拒染法判断,新鲜分离的肝细胞平均活力为95%。然后将分离的肝实质细胞在威斯康星大学溶液(UW)中于0℃保存长达1周。在这种低温保存期间,0℃下1天后活力仅略有下降至92%,3天后降至85%。此后,活力迅速下降。冷藏长达3天后,可以将肝实质细胞用于短期悬浮培养或细胞培养。新鲜分离的肝细胞和冷藏2天后的肝细胞在细胞培养中的贴壁效率相同(84%)。新鲜分离的细胞与低温保存3天后的细胞相比,细胞色素P450含量以及可溶性环氧化物水解酶、尿苷二磷酸葡萄糖醛酸基转移酶、酚磺基转移酶和谷胱甘肽S-转移酶的酶活性无显著差异。此外,将新鲜分离的和保存3天的完整肝细胞用作细胞介导的沙门氏菌致突变性试验中的代谢系统。新鲜的和保存的肝实质细胞均将苯并(a)芘、2-氨基蒽和环磷酰胺代谢为其最终诱变剂。(摘要截短于250字)

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