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随着国内新冠病毒肺炎疫情的持续向好,我们已经从“抗疫”转变为“防疫”状态,而与此同时,国外疫情却还处于爆发阶段。为了控制SARS-CoV-2 (2019-nCoV),无数科研工作者就开始了争分夺秒的科研攻关,围绕新冠病毒做了大量的研究。

3分pk10为助力新型冠状病毒的相关研究和检测,普诺赛(Procell)总结提炼了新型冠状病毒研究相关的细胞产品,并将细胞相关的参考信息汇总如下,希望能为广大科研工作者带来一点帮助。

主要包括以下内容:

*欢迎各位老师提供更多相关的附加信息、更正、更新,我们将及时更新完善。联系邮箱:techsupport@sadiesoap.com

已被证实可用于扩增SARS-CoV-2的细胞

1.Vero 系列

以下3个“Vero”系列的细胞系被广泛应用于SARS-CoV-2的培养扩增(按照SARS-CoV-2增殖效果递减顺序展示)。

1.1 Vero C1008(更常被称为Vero E6,细胞系产品推荐:CL-0491)

参考文献:

  • [1]. Matsuyama et al. (PubMed=32165541): "However, the viral RNA copies in the VeroE6/TMPRSS2 cell culture supernatants were >100 times greater than those from VeroE6 cells."

  • [2]. Harcourt et al. (DOI=10.1101/2020.03.02.972935): "We found that the virus replicates to high titer in Vero-CCL81 cells and Vero E6 cells in the absence of trypsin." (also see figure 3A)

  • [3]. Wang et al. (PubMed=32020029): "Vero E6 cells were infected with nCoV-2019BetaCoV/Wuhan/WIV04/2019 at a multiplicity of infection (MOI) of 0.05 in the presence of varying concentrations of the test drugs."

  • [4]. Zhu et al. (PubMed=31978945): "Virus isolation from the clinical specimens was performed with human airway epithelial cells and Vero E6 and Huh-7 cell lines" and "No specific cytopathic effects were observed in the Vero E6 and Huh-7 cell lines until 6 days after inoculation."

  • [5]. Lokugamage et al. (DOI=10.1101/2020.03.07.982264): "Using Vero E6 cells, we demonstrate that SARS-CoV-2 maintains similar viral replication kinetics as SARS-CoV following a low dose infection."

  • [6]. Kim et al. (PubMed=32149036): "The SARS-CoV-2 could replicate in other cells (Vero E6 and Caco-II cells), in addition to Vero cells (data not shown)."

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1.2 Vero (细胞系产品推荐:CL-0242)

参考文献:

  • [1]. Harcourt et al. (DOI=10.1101/2020.03.02.972935): "We found that the virus replicates to high titer in Vero-CCL81 cells and Vero E6 cells in the absence of trypsin." (also see figure 3A)

  • [2]. Matsuyama et al. (PubMed=32165541): "the amount of SARS-CoV-2 RNAs in the culture supernatants of Vero, Calu-3, and A549 cells 48 h p.i. was low."

  • [3]. Sheahan et al. (DOI=10.1101/2020.03.19.997890): "NHC strongly inhibited SARS-CoV-2 replication in Vero cells with an IC50 of 0.3muM and CC50 of >10muM."

  • [4]. Kim et al. (PubMed=32149036): "The virus replicated in Vero cells and cytopathic effects were observed."

  • [5]. Ou et al. (PubMed=32221306): "Huh7 and Vero 81 cells also gave about 10-fold increase in luciferase activities when transduced by SARS-CoV-2."

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1.3 VeroE6/TMPRSS2

参考文献:

Matsuyama et al. (PubMed=32165541): "VeroE6/TMPRSS2 cells are superior to other cell lines tested in this study for SARS-CoV-2 isolation."


2.其他系列

2.1 Caco-2 (细胞系产品推荐:CL-0050)

参考文献:

According to Kim et al. (PubMed=32149036): "The SARS-CoV-2 could replicate in other cells (Vero E6 and Caco-II cells), in addition to Vero cells (data not shown)."


2.2 Calu-3 (细胞系产品推荐:CL-0054)

参考文献:

  • [1]. According to Matsuyama et al. (PubMed=32165541): "the amount of SARS-CoV-2 RNAs in the culture supernatants of Vero, Calu-3, and A549 cells 48 h p.i. was low."

  • [2]. According to Ou et al. (PubMed=32221306): "Calu3 is highly susceptible to SARS-CoV-2-S-mediated entry" and "Compared to mock control, SARS-CoV-2 S pseudovirions showed an over 500-fold increase in luciferase activities in Calu3 cells, at a level similar to SARS-CoV S pseudovirions."Note: UTMB的Tseng团队于2010年曾经发表过(PubMed=20090954) Calu-3 2B4(Calu-3的克隆),该细胞系ACE2表达增加,并且对SARS冠状病毒(SARS-CoV)感染易感性高。

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2.3 HEK293T (细胞系产品推荐:CL-0005)

参考文献:

Harcourt et al. (DOI=10.1101/2020.03.02.972935): "both HUH7.0 and 293T cells showed only modest viral replication" (also see figure 3A).


2.4 Huh-7 (细胞系产品推荐:CL-0120)

参考文献:

  • [1]. According to Harcourt et al. (DOI=10.1101/2020.03.02.972935): "both HUH7.0 and 293T cells showed only modest viral replication" (also see figure 3A).

  • [2]. According to Zhu et al. (PubMed=31978945): "Virus isolation from the clinical specimens was performed with human airway epithelial cells and Vero E6 and Huh-7 cell lines" and "No specific cytopathic effects were observed in the Vero E6 and Huh-7 cell lines until 6 days after inoculation."

  • [3]. According to Ou et al. (PubMed=32221306): "Huh7 and Vero 81 cells also gave about 10-fold increase in luciferase activities when transduced by SARS-CoV-2."

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2.5 LLC-MK2 (细胞系产品推荐:CL-0141)

参考文献:

Ou et al. (PubMed=32221306): "LLCMK2 cells, a rhesus monkey kidney epithelium cell line, exhibited different susceptibility to SARS-CoV S and SARS-CoV-2 S transduction, but the reasons for this are currently not known and require further investigation."

已被证实不适合扩增SARS-CoV-2的细胞系

A-549 (细胞系产品推荐:CL-0016)

参考文献:

  • [1]. According to Matsuyama et al. (PubMed=32165541): "the amount of SARS-CoV-2 RNAs in the culture supernatants of Vero, Calu-3, and A549 cells 48 h p.i. was low."

  • [2]. According to Harcourt et al. (DOI=10.1101/2020.03.02.972935): "A549 cells were incompatible with SARS-CoV-2 infection" (also see figure 3A).

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Efk3B from bat

参考文献:

Harcourt et al. (DOI=10.1101/2020.03.02.972935): "In addition, SARS-CoV-2 failed to replicate in the bat EFK3B cells which are susceptible to MERS-CoV." (also see figure 3A).

基于细胞宿主的疫苗相关细胞系

一些公司和研究机构正在全力研发基于细胞的SARS-CoV-2疫苗。以下细胞系是这类研究中需要用到的:

CHO-K1(细胞系产品推荐:CL-0062) 或其衍生物中的一种可被用于CSIRO: (http://www.theage.com.au/national/coronavirus-outbreak-how-the-covid-19-vaccine-is-being-made-20200220-p542rh.html)"On February 21 the team's vaccine draft was sent to the CSIRO's manufacturing facility in the Melbourne suburb of Clayton, and manufacturing of a pilot dose began. The facility houses a 200-litre fermenting vat that will be filled with a "soup" of Chinese hamster ovary cells - the best cells for the task - in a nutritious goo."

Sorrento 使用的是K-562(细胞系产品推荐:CL-0130)Therapeutics, Inc.(DOI=http://doi.org/10.1016/j.medidd.2020.100026): see Ji et al: "Engineered Spike-1 protein is expressed on the surface of K562 human myelogenous leukemia cells via introduction of expression constructs into the cellular genome allowing for stable expression of the transgene. Stable-modified K562 clones are selected, profiled for Spike-1 expression as well as overall immunogenic potency, and prepared as GMP master cell bank to be used for large scale manufacturing. Irradiated cells are formulated as vaccine product and administered via intramuscular or subcutaneous injection."

PER.C(http://www.jnj.com/coronavirus): "Johnson & Johnson is mobilizing resources of its Janssen Pharmaceutical Companies in response to the outbreak to develop a possible preventive vaccine candidate against SARS-CoV-2, leveraging Janssen's AdVac (R) and PER.C6 (R) technology, that provide the ability to rapidly upscale production of the optimal vaccine candidate."