In vitro EBV Infection of Mononuclear Cells that Have Been Cryo-preserved
冷藏单核细胞的体外艾普斯登巴尔病毒（EBV）感染试验
发布: 2014年07月20日第4卷第14期 DOI: 10.21769/BioProtoc.1177 浏览次数: 7708
评审: Anonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Dec 2013
Abstract
Epstein-Barr Virus (EBV) is a B-lymphotropic herpesvirus which the majority of adult human population is latently-infected with. Various immunological and molecular in vitro studies have been facilitated by the use of EBV’s ability to infect and transform B cells to immortalized polyclonal B cell lines. Many of these studies use freshly isolated cord-blood mononuclear cells (CBMC). Some experiments may, however, require EBV infection of samples that have been prospectively collected and cryo-preserved. Here we share a protocol that we used to successfully infect B cells from cryo-preserved CBMCs and peripheral-blood mononuclear cells (PBMC) (Sohlberg et al., 2013; Saghafian-Hedengren et al., 2013).
Materials and Reagents
- Cryo-preserved CBMC
- Cryo-preserved PBMC
- EBV strain B95-8 containing supernatant
Note: The batch here had a titer of 2.5 x 105 Ramos infectious units. RaIU, which was determined by infection of the EBV negative Burkitts lymphoma B-cell line Ramos, followed by anti-complement immunoflourescent assay (ACIF) to detect the number of infected cells.
- RPMI 1640 (Life Technologies)
- 10% heat-inactivated fetal-calf serum (Hyclone)
- L-glutamine (2 mmol/l)
- Penicillin G-sodium (100 U/ml)
- Streptomycin sulfate (100 mg/ml) (Merck KGaA)
- Complete cell-culture medium (see Recipes)
Equipment
- 48-well flat-bottomed tissue-culture treated plates (Sarstedt AG)
- Humified incubator with for adjustment of 37 °C and 5% CO2 for cell culture
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Saghafian-Hedengren, S., Sohlberg, E., Theorell, J., Carvalho-Queiroz, C., Nagy, N., Persson, J., Nilsson, C., Bryceson, Y. T. and Sverremark-Ekström, E. (2014). In vitro EBV Infection of Mononuclear Cells that Have Been Cryo-preserved. Bio-protocol 4(14): e1177. DOI: 10.21769/BioProtoc.1177.
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分类
微生物学 > 微生物-宿主相互作用 > 病毒
微生物学 > 微生物细胞生物学 > 细胞分离和培养
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