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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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


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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