在模拟微重力条件下培养的HeLa细胞中鼻病毒的复制

Rhinovirus replication in HeLa cells cultured under conditions of simulated microgravity.

作者信息

Long J P, Pierson S, Hughes J H

机构信息

Children's Hospital Research Foundation, Columbus, OH 43205, USA.

出版信息

Aviat Space Environ Med. 1998 Sep;69(9):851-6.

PMID:9737755

Abstract

BACKGROUND

Rotating-wall vessels (RWVs) allow for the growth of cells under conditions of simulated microgravity. Information about the replication of viruses in simulated microgravity using RWVs has not been reported. Cells grown in RWVs are subjected to low shear motion, and the replication of certain viruses such as rhinoviruses has been reported to be enhanced by motion.

HYPOTHESIS

Our research was based on the hypothesis that rhinovirus replication would be enhanced under conditions of simulated microgravity.

METHODS

HeLa cells were cultured in three-dimensional cultures on microcarrier beads in simulated microgravity using RWVs and in sealed Teflon roller bottles. Two-dimensional cultures of HeLa cells were also grown in tissue culture flasks (T-150s). Viral infections for all cultures were carried out under standardized conditions at 1 x g. The amount of new virus released during the first viral replication cycle and the total viral yields obtained from multiple viral replication cycles were determined.

RESULTS

Viral quantitation during the first viral replication cycle showed that after 10-13 h RWV and Teflon roller bottle supernatants contained significantly more virus than the supernatants from T-150 cultures. After multiple viral replication cycles (at 24, 48, 72, and 96 h following infection), total viral samples (both free and cell-associated virus) from RWV cultures contained significantly more virus than Teflon roller bottle cultures.

CONCLUSIONS

The rhinovirus replication cycle was enhanced in cultures grown in the presence of motion (Teflon roller bottle cultures and RWV cultures). Additionally, multiple rounds of rhinovirus replication yielded more virus in simulated microgravity conditions. Viral transmission in cell cultures in RWVs was efficient and was similar to or better than what occurred in the Teflon roller bottles. The cultivation of cells in simulated microgravity possibly affected the rate of viral adsorption/uptake, the viral replication cycle, and/or the viral yield. RWVs provide an effective means for culturing human rhinoviruses.

摘要

背景

旋转壁式生物反应器（RWV）可使细胞在模拟微重力条件下生长。关于利用RWV在模拟微重力条件下病毒复制的信息尚未见报道。在RWV中生长的细胞会受到低剪切力作用，且据报道某些病毒（如鼻病毒）的复制会因运动而增强。

假设

我们的研究基于这样的假设，即鼻病毒在模拟微重力条件下的复制会增强。

方法

使用RWV在模拟微重力条件下，将HeLa细胞在微载体珠上进行三维培养，并在密封的特氟龙滚瓶中培养。HeLa细胞的二维培养也在组织培养瓶（T - 150）中进行。所有培养物的病毒感染均在1×g的标准化条件下进行。测定了第一个病毒复制周期中释放的新病毒量以及多个病毒复制周期获得的总病毒产量。

结果

第一个病毒复制周期的病毒定量显示，在10 - 13小时后，RWV和特氟龙滚瓶上清液中的病毒含量明显高于T - 150培养物的上清液。在多个病毒复制周期后（感染后24、48、72和96小时），RWV培养物的总病毒样本（游离病毒和细胞相关病毒）中的病毒含量明显高于特氟龙滚瓶培养物。

结论

在有运动的培养物（特氟龙滚瓶培养物和RWV培养物）中，鼻病毒复制周期增强。此外，在模拟微重力条件下，多轮鼻病毒复制产生了更多病毒。RWV中细胞培养物中的病毒传播效率高，与特氟龙滚瓶中的情况相似或更好。在模拟微重力条件下培养细胞可能影响病毒吸附/摄取速率、病毒复制周期和/或病毒产量。RWV为培养人鼻病毒提供了一种有效手段。