Oncotarget

Research Papers:

Establishment of mouse neuron and microglial cell co-cultured models and its action mechanism

Bo Zhang, Yunfeng Yang, Jun Tang, Yihao Tao, Bing Jiang, Zhi Chen, Hua Feng, Liming Yang _ and Gang Zhu

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Oncotarget. 2017; 8:43061-43067. https://doi.org/10.18632/oncotarget.17898

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Abstract

Bo Zhang1, Yunfeng Yang1, Jun Tang1, Yihao Tao1, Bing Jiang1, Zhi Chen1, Hua Feng1, Liming Yang1 and Gang Zhu1

1Department of Neurosurgery, Southwest Hospital,Third Military Medical University, Chongqing, China

Correspondence to:

Liming Yang, email: yangliming212@163.com

Gang Zhu, email: gangzhu6666@sina.com

Keywords: neuron, microglial cell, OGD, tOGD

Received: February 12, 2017     Accepted: April 15, 2017     Published: May 16, 2017

ABSTRACT

Objective: The objective of this study is to establish a co-culture model of mouse neurons and microglial cells, and to analyze the mechanism of action of oxygen glucose deprivation (OGD) and transient oxygen glucose deprivation (tOGD) preconditioning cell models.

Results: Mouse primary neurons and BV2 microglial cells were successfully cultured, and the OGD and tOGD models were also established. In the co-culture of mouse primary neurons and microglial cells, the cell number of tOGD mouse neurons and microglial cells was larger than the OGD cell number, observed by a microscope. CCK-8 assay result showed that at 1h after treatment, the OD value in the control group is lower compared to all the other three groups (P < 0.05). The treatment group exhibited the highest OD value among the four groups. The results observed at 5h were consistent with the results at 1 h. Flow cytometry results showed that at 1h after treatment the apoptosis percentages is higher in the control group compared to other three groups (P < 0.05).

Materials and Methods: Mouse brain tissues were collected and primary neurons cells were cultured. In the meantime mouse BV2 microglia cells were cultured. Two types of cells were co-cultured, and OGD and tOGD cell models were established. There were four groups in the experiment: control group (OGD), treatment group (tOGD+OGD), placebo group (tOGD+OGD+saline) and minocycline intervention group (tOGD+OGD+minocycline). CCK-8 kit was used to detect cell viability and flow cytometry was used to detect apoptosis.

Conclusions: In this study, mouse primary neurons and microglial cells were co-cultured. The OGD and tOGD models were established successfully. tOGD was able to effectively protect neurons and microglial cells from damage, and inhibit the apoptosis caused by oxygen glucose deprivation.


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