Zhang Bo, Yang Yunfeng, Tang Jun, Tao Yihao, Jiang Bing, Chen Zhi, Feng Hua, Yang Liming, Zhu Gang
Department of Neurosurgery, Southwest Hospital,Third Military Medical University, Chongqing, China.
Oncotarget. 2017 Jun 27;8(26):43061-43067. doi: 10.18632/oncotarget.17898.
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.
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).
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.
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.
本研究旨在建立小鼠神经元与小胶质细胞的共培养模型，并分析氧糖剥夺（OGD）和短暂氧糖剥夺（tOGD）预处理细胞模型的作用机制。
成功培养了小鼠原代神经元和BV2小胶质细胞，并建立了OGD和tOGD模型。在小鼠原代神经元与小胶质细胞的共培养中，通过显微镜观察发现，tOGD小鼠神经元和小胶质细胞的细胞数量多于OGD细胞数量。CCK-8检测结果显示，处理后1小时，对照组的OD值低于其他三组（P<0.05）。治疗组在四组中OD值最高。5小时观察结果与1小时结果一致。流式细胞术结果显示，处理后1小时，对照组的凋亡率高于其他三组（P<0.05）。
采集小鼠脑组织并培养原代神经元细胞。同时培养小鼠BV2小胶质细胞。将两种细胞进行共培养，并建立OGD和tOGD细胞模型。实验分为四组：对照组（OGD）、治疗组（tOGD+OGD）、安慰剂组（tOGD+OGD+生理盐水）和米诺环素干预组（tOGD+OGD+米诺环素）。使用CCK-8试剂盒检测细胞活力，流式细胞术检测细胞凋亡。
本研究将小鼠原代神经元与小胶质细胞进行了共培养。成功建立了OGD和tOGD模型。tOGD能够有效保护神经元和小胶质细胞免受损伤，并抑制氧糖剥夺引起的细胞凋亡。
