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[细胞培养控制中的几个方法学问题]

[Several methodologic problems in the control of cell cultures].

作者信息

Demidova S A, Tsareva A A, mikhaĭlova G R, Perekrest V V, Gushchin B V

出版信息

Vopr Virusol. 1976 May-Jun(3):371-9.

PMID:983006
Abstract

Some human and animal continuous cell lines as well as primary cell cultures were examined by karyological, electron microscopial, virological and molecular biological methods and also by the electrophoretic motility of glucose-6-phosphate dehydrogenase (G-6-PDG) in polyacrylamide gel. All human and animal continuous cell lines were shown to contain mycoplasma, 17-to contain intracytoplasmic particles of type A oncornaviruses, 5 -- type B oncornaviruses similar to Mason-Pfizer virus, 8 -- paramyxoviruses, 2 --oncornaviruses type C. A high molecular RNA with sedimentation constant 64--70 S was found in oncornaviruses isolated from cell cultures. Intracellular virus or subviral structures were detected by association of the reverse transcriptase activity with high molecular RNA. The presence in the cell cultures of marker chromosomes of HeLa cells, the absence in these cultures of Y chromosomes, the presence of the G-6-PDG enzyme with type A motility indicate the possibility of contamination of human continuous cell lines with HeLa cells.

摘要

采用染色体学、电子显微镜学、病毒学和分子生物学方法,以及通过葡萄糖-6-磷酸脱氢酶(G-6-PDG)在聚丙烯酰胺凝胶中的电泳迁移率,对一些人和动物的连续细胞系以及原代细胞培养物进行了检测。结果表明,所有的人和动物连续细胞系均含有支原体,17个含有A型肿瘤病毒的胞质内颗粒,5个含有类似于梅森-辉瑞病毒的B型肿瘤病毒,8个含有副粘病毒,2个含有C型肿瘤病毒。从细胞培养物中分离出的肿瘤病毒中发现了沉降常数为64-70S的高分子量RNA。通过逆转录酶活性与高分子量RNA的关联检测到细胞内病毒或亚病毒结构。细胞培养物中存在HeLa细胞的标记染色体,这些培养物中不存在Y染色体,存在具有A型迁移率的G-6-PDG酶,表明人连续细胞系有可能被HeLa细胞污染。

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