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一种人T淋巴母细胞系的异质性。

Heterogeneity of a human T-lymphoblastoid cell line.

作者信息

Snow K, Judd W

出版信息

Exp Cell Res. 1987 Aug;171(2):389-403. doi: 10.1016/0014-4827(87)90171-6.

Abstract

A human T-lymphoblastoid cell line (Jurkat) was cloned, and four resulting sublines were characterized in a variety of ways with the objective of gaining information on heterogeneity in cell lines. Within a few weeks of cloning, distinct cellular morphologies and growth patterns became apparent in the four sublines. Growth rate measurements made over 3 months did not show any significant differences between the sublines. Surface protein profiles obtained by radioimmunoprecipitation using antisera in conjunction with extracts from [35S]Met and 125I-labeled cells revealed differences between the sublines. Analysis of total cell DNA showed that one of the sublines possessed only half the chromosome complement of the other sublines and the parental line. Karyotyping confirmed this result and, in addition, demonstrated that chromosome numbers fluctuated around a mean value for each subline. Karyotypic variability became apparent within 2 months of cloning and tended to increase with time in culture. G-banding analysis showed that the analyzed cell populations contained distinctive cytogenetic aberrations. Properties of the cloned sublines were monitored over a 9-month period. One of the sublines that had shown heterogeneous morphology even after 6 weeks maintained the heterogeneity throughout this time. Another subline underwent a marked change in morphology (round to irregular) and growth habit (single cells to large clumps) with increasing time in culture. Interestingly, several alterations to surface proteins accompanied these growth changes. A third subline had relatively stable morphology and chromosome number throughout the 9-month period. The modal chromosome number was hypotetraploid for three sublines and the parent line, but was diploid for another subline. However, it was interesting that progression toward tetraploidy in this subline was apparent after almost 2 years of culturing. The results showed that the original cell line consisted of a heterogeneous assemblage of cell types, some of which were quite unstable. Some implications for research using cultured cell lines are discussed.

摘要

克隆了一株人T淋巴细胞样细胞系(Jurkat),并通过多种方式对由此产生的四个亚系进行了表征,目的是获取有关细胞系异质性的信息。在克隆后的几周内,四个亚系中出现了明显不同的细胞形态和生长模式。在3个月内进行的生长速率测量未显示各亚系之间有任何显著差异。使用抗血清结合来自[35S]甲硫氨酸和125I标记细胞的提取物进行放射免疫沉淀获得的表面蛋白谱显示各亚系之间存在差异。对总细胞DNA的分析表明,其中一个亚系的染色体组仅为其他亚系和亲本系的一半。核型分析证实了这一结果,此外还表明每个亚系的染色体数围绕一个平均值波动。核型变异性在克隆后2个月内变得明显,并随着培养时间的延长而趋于增加。G带分析表明,所分析的细胞群体包含独特的细胞遗传学异常。在9个月的时间里监测克隆亚系的特性。其中一个亚系即使在6周后仍表现出异质形态,在此期间一直保持着异质性。另一个亚系随着培养时间的增加,形态(圆形变为不规则形)和生长习性(单细胞变为大团块)发生了显著变化。有趣的是,这些生长变化伴随着表面蛋白的一些改变。第三个亚系在整个9个月期间具有相对稳定的形态和染色体数。三个亚系和亲本系的众数染色体数为亚四倍体,但另一个亚系为二倍体。然而,有趣的是,在培养近2年后,这个亚系向四倍体的进展变得明显。结果表明,原始细胞系由不同类型的细胞组成,其中一些细胞相当不稳定。讨论了对使用培养细胞系进行研究的一些启示。

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