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在遗传研究中应用爱泼斯坦-巴尔病毒优化永生化B淋巴细胞的产生作为阳性对照

Application of Epstein-Barr Virus for Optimization of Immortalized B-lymphocyte Production as a Positive Control in Genetic Studies.

作者信息

Tousizadeh Behnaz, Moghim Sharareh, Chaleshtori Ahmad Reza Salehi, Ghanbarian Maryam, Mirian Mina, Salehi Mansoor, Tousizadeh Sepideh, Zaboli Fatemeh

机构信息

Department of Microbiology, Faculty of Basic Sciences, Ayatollah Amoli Azad University, Amol, Iran.

Pediatric Inherited Disease Research Center, Isfahan University of Medical Sciences, Isfahan, Iran.

出版信息

Adv Biomed Res. 2017 Jul 14;6:80. doi: 10.4103/2277-9175.210659. eCollection 2017.

Abstract

BACKGROUND

Infection of B-cells with Epstein-Barr virus (EBV) leads to more and subsequent immortalization. This is considered as the method of choice for generating lymphoblastoid cell lines (LCLs). Producing LCLs, although very useful but is very time consuming and troublesome, drives the requirement for quicker and more reliable methods for EBV-driven B-cell transformation.

MATERIALS AND METHODS

After successfully production of LCLs, different parameters including temperature, serum concentration, type of culture medium, and CO concentration were evaluated on EBV-transformed B-cells. In this study, we were able to produce LCLs and optimize condition.

RESULTS

The best condition for generating LCLs was 37°C, 5% CO, 20% fasting blood sugar, and RPMI 1640. The study results were to establish a reliable method for producing LCLs that can be used to produce immortalized B-cells from almost any sources.

CONCLUSION

This can help with tumorgenecity studies, as well as producing control material for rare genetic disorders and so on. The aim of this study was to determine optimized condition for reliable and reproducible LCLs from different sources.

摘要

背景

爱泼斯坦-巴尔病毒(EBV)感染B细胞会导致更多细胞随后发生永生化。这被认为是生成淋巴母细胞系(LCLs)的首选方法。尽管生成LCLs非常有用,但非常耗时且麻烦,这促使人们需要更快、更可靠的EBV驱动的B细胞转化方法。

材料与方法

成功生成LCLs后,对EBV转化的B细胞评估了包括温度、血清浓度、培养基类型和二氧化碳浓度等不同参数。在本研究中,我们成功生成了LCLs并优化了条件。

结果

生成LCLs的最佳条件是37°C、5%二氧化碳、20%空腹血糖和RPMI 1640培养基。研究结果是建立了一种可靠的生成LCLs的方法,可用于从几乎任何来源生成永生化B细胞。

结论

这有助于肿瘤发生研究,以及为罕见遗传疾病等生产对照材料。本研究的目的是确定从不同来源可靠且可重复地生成LCLs的优化条件。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/54a4/5539668/fda8e330a60f/ABR-6-80-g001.jpg

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