细胞描述	本库的细胞仅用于科研工作，未经许可不得用于其他目的，使用者不得将本库细胞转让给第三者。

 细胞收到后处理：

细胞在培养瓶中培养至良好状态后灌满完全培养液并封好瓶口是细胞运输的最好办法。收到细胞回到自己的实验室后，先打开外包装，用75%酒精喷洒整个瓶消毒后放到超净台内，严格无菌操作，培养箱静置2-4小时。镜下观察：未超过80%汇合度时，可将瓶装的完全培养液收集至离心管中，加入6ml完全培养基，放入37℃、5%CO2孵箱培养；超过80%汇合度时，根据情况传代或者冻存，具体操作见细胞培养步骤。（注意发货的是密封培养瓶的话，放入培养箱培养记得培养瓶盖子拧松）

细胞培养步骤：

1）复苏细胞：将含有1mL细胞悬液的冻存管在37℃水浴中迅速摇晃解冻，加入5mL培养基混合均匀。在1000RPM条件下离心5分钟，弃去上清液，补加4-6mL完全培养基后吹匀。然后将所有细胞悬液加入培养瓶中培养过夜（或将细胞悬液加入6cm皿中），培养过夜。第二天换液并检查细胞密度。

2）细胞传代：如果细胞密度达80%-90%，即可进行传代培养。

1、对于贴壁细胞，传代可参考以下方法：

1. 弃去培养上清，用不含钙、镁离子的PBS润洗细胞1-2次。

2. 加1-2ml消化液（0.25%Trypsin-0.53mM EDTA）于培养瓶中，置于37℃培养箱中消化1-2min，然后在显微镜下观察细胞消化情况，若细胞大部分变圆并脱落，迅速拿回操作台，轻敲几下培养瓶后加5ml以上含10%血清的完全培养基终止消化。

3.轻轻吹打细胞，完全脱落后吸出，在1000RPM条件下离心8-10分钟，弃去上清液，补加1-2mL培养液后吹匀。

4. 按5-6ml/瓶补加培养液，将细胞悬液按1：2到1：5的比例分到新的含5-6 ml培养液的新皿中或者瓶中。

PS：若客户收到2ml小管细胞，收到细胞后，用75%酒精喷洒整个管子消毒后放到超净台或安全柜内，严格无菌操作；将小管细胞转移至T25培养瓶或6cm培养皿，加入5ml左右完全培养基混匀，放入培养箱过夜培养后查看细胞密度：若密度未超过80%，换液继续培养，视情况传代或者冻存。若密度超过80%，可直接进行传代（方法同上）。

文献参考： 文章标题：Salinomycin inhibits orthopoxvirus infection in vitro and in vivo by blocking endosomal acidification

作者列表：Congcong Zhang, Yanhua He, Jiaqi Wang, Wanda Tang, Dawei Wang, Zhendong Pan, Nuo Liu, Xu Zheng, Hailin Tang, Zhongtian Qi, Binghui Xia, Ping Zhao

影响因子：4

期刊：ANTIVIRAL RESEARCH

发表时间：2026-4-4

DOI:10.1016/j.antiviral.2026.106403

文献主题：Abstract

The recent global spread of monkeypox virus (MPXV) highlights the urgent need for effective antiviral therapies. Through high-throughput screening of an FDA-approved small molecule drug library, we identified salinomycin as a potent inhibitor of orthopoxvirus infection. Salinomycin showed nanomolar anti-MPXV activity in human keratinocytes (HaCaT) and lung epithelial cells (A549), with selectivity indices >100. Importantly, its efficacy was validated in primary human fibroblasts, reducing infectious viral titers by ∼2.6 log units (440-fold) versus controls. Mechanistic studies revealed salinomycin acts at the viral post-entry stage, blocking membrane fusion via disrupting endosomal acidification, without affecting clathrin-mediated endocytosis. This host-directed mechanism was validated by DiO fusion assays, acridine orange staining, and low-pH pulse rescue experiments. In a lethal intranasal vaccinia virus (VACV-WR) murine challenge model, oral salinomycin (1 mg/kg/day) significantly reduced lung viral loads, mitigated histopathology, and conferred full mortality protection. Furthermore, salinomycin synergized with tecovirimat, an approved viral egress inhibitor. In fixed-ratio combinations, salinomycin's EC50 dropped ∼4.0-fold (30.12 nM to 7.50 nM), and tecovirimat's ∼3.2-fold (11.96 nM to 3.73 nM). Bliss independence analysis confirmed strong synergy, with a maximum ΔBliss value of 16.97. Notably, the combination achieved 100% survival in a high-dose VACV-WR model, an effect unmatched by either monotherapy. Collectively, these preliminary in vitro and in vivo data identify salinomycin as a potential host-targeted anti-orthopoxvirus inhibitor, especially in combination with tecovirimat. Further comprehensive preclinical evaluations, including full toxicological profiling and validation in clinically relevant models, are needed to define its translational potential for MPXV and other orthopoxvirus infections.

文献链接：

Salinomycin inhibits orthopoxvirus infection in vitro and in vivo by blocking endosomal acidification - ScienceDirect