体外呼吸道上皮细胞细菌黏附模型的建立

王若茜; 枉前; 孙凤军; 刘耀; 张镭; 李晓宇; 夏培元

doi:10.16016/j.1000-5404.201802030

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体外呼吸道上皮细胞细菌黏附模型的建立

基础医学 | 更新时间：2026-01-09

- 体外呼吸道上皮细胞细菌黏附模型的建立
- Establishment of an in vitro model of bacteria adhesion to respiratory epithelial cells
- 第三军医大学学报 2018年40卷第11期页码：973-978
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金面上项目(81373451)
- DOI：10.16016/j.1000-5404.201802030
  中图分类号：
- 收稿：2018-02-05，
  
  纸质出版：2018-06
- 稿件说明：
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王若茜, 枉前, 孙凤军, 等. 体外呼吸道上皮细胞细菌黏附模型的建立[J]. 第三军医大学学报, 2018,40(11):973-978.

Ruoxi WANG, Qian WANG, Fengjun SUN, et al. Establishment of an in vitro model of bacteria adhesion to respiratory epithelial cells[J]. Journal of Third Military Medical University, 2018, 40(11): 973-978.
王若茜, 枉前, 孙凤军, 等. 体外呼吸道上皮细胞细菌黏附模型的建立[J]. 第三军医大学学报, 2018,40(11):973-978. DOI： 10.16016/j.1000-5404.201802030.

Ruoxi WANG, Qian WANG, Fengjun SUN, et al. Establishment of an in vitro model of bacteria adhesion to respiratory epithelial cells[J]. Journal of Third Military Medical University, 2018, 40(11): 973-978. DOI： 10.16016/j.1000-5404.201802030.

摘要

Abstract

Objective

To establish an

in vitro

model to study the adhesion of bacteria to respiratory epithelial cells based on the construction of flow-chamber.

Methods

After 1×10

HBE cells were seeded in the flow chamber pre-coated with 2 mg/mL bovine collagen

the chamber was cultured in RPMI 1640 medium containing 20% serum and incubated at 37℃ in a 5% CO

incubator for 24 h. Once the bottom was covered

with HBE cells

the chamber could be used for further experiments. When the number of cells in the flow chamber was taken as the evaluation index

the effects of the flow rate

the flow duration and the mobile phase composition on the cellular model were investigated by orthogonal design

and the optimal experimental conditions were screened. Then under the obtain optimal conditions

after the bacteria were adhered to the cells at 2×10

5×10

and 2.5×10

CFU/mL inoculated concentrations respectively

the amount of bacterial adhesion was taken as an index

and the results were compared with the conventional plate culture for bacterial adhering to cells

so as to determine whether the flow chamber is a viable model of bacteria adhesion to cells. The amount of bacteria adhering to cells was characterized by SYTO9 fluorescently labeled bacteria.

Results

The influencing factors in optimal experimental conditions in order of their impacts were as follows:flow rate

flow duration

mobile phase composition. The ANOVA study showed that the impacts of flow rate and flow duration were significant (

0.05)

and while those of mobile phase composition were not. After fluorescent staining

the model realized the real-time fluorescence observation of the bacteria adhesion to the respiratory epithelial cells. According to the quantitative results of bacterial adhesion

with the increase of multiplicity of infection

both the flow chamber and the conventional culture plate detected the elevated amount of adhering bacteria

in a linear manner. However

with the same multiplicity of infection

the conventional culture plate model detected significantly larger amount of adhering bacteria when compared to the flow chamber (

0.05).

Conclusion

Under appropriate flow rate and flow duration

our model is an alternative to the conventional plate for studying the adhesion of bact

eria to the airway epithelial cells

with the advantages of being more close to the

in vivo

environment

high accuracy and real-time observation.

关键词

Keywords

references

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