摘要
目的采用FX-4000TTM柔性基底加载系统建立体外大鼠脊髓源性少突胶质前体细胞机械拉伸损伤模型,为进一步探讨少突胶质前体细胞在脊髓损伤及修复过程中的作用及机制提供模型基础。方法采用振摇法体外纯化培养大鼠脊髓少突胶质前体细胞,并进行免疫荧光细胞鉴定。将纯化培养3 d的少突胶质前体细胞随机分为A组(对照组)、B组(拉伸强度5%)、C组(拉伸强度10%)、D组(拉伸强度15%)。采用FX-4000TTM柔性基底加载系统机械拉伸少突胶质前体细胞2 h后,通过倒置显微镜观察各组细胞形态学改变、MTT法检测细胞存活率、双染流式细胞术检测细胞凋亡,对损伤模型进行评价。结果体外成功培育少突胶质前体细胞且纯度>90%。B组拉伸强度对细胞形态及存活没有明显影响,基本不构成损伤。C组和D组细胞存活率较A组显著降低(P<0.05),凋亡率也明显升高(P<0.05),并出现明显病理性形态改变;但D组有明显的细胞脱落现象且细胞存活率降到<50%,不利于后续实验进行。结论采用FX-4000TTM柔性基底加载系统以10%为拉伸强度可建立有效的少突胶质前体细胞机械拉伸损伤模型。
Objective To establish a model of stretch-induced mechanical injury on cultured oligodendrocyte precursor cells （OPCs） in vitro with the FX-4000TTM system,in order to further explore the role and mechanism of OPCs in spinal cord injury and repair.Methods OPCs were separated from spinal cord of neonatal rats and then purified with the shaking method in vitro.Immunofluorescence assay was applied to identify the separated cells.A FX-4000TM system was used to apply cyclic stretch.The purified OPCs were randomly divided into four groups：group A （control group）,group B （5% tensile strain）,group C （10% tensile strain）,group D （15% tensile strain）.After OPCs were strained for 2 h by the FX-40OOTTM system,morphology of OPCs was observed by using inverted microscope,viability of OPCs was measured by MTT assay and apoptosis of the cells was detected by double staining flow cytometry,damage models were evaluated. Results The percentage of purified OPCs in vitro was more than 90%.There were no significant differences between group A and group B in survival rate and cell morphology.Compared with group A,survival rate of OPCs in group C and D was significantly decreased （P〈0.05）,apoptosis rate of OPCs was significantly increased （P〈0.05）,and obvious patho- logical morphologic changes of OPCs were also observed in group C and D.However,there was a large loss of cell attach- ment and survival rate of OPCs was found to be less than 50% in group D.Conelusion The model of stretch-induced mechanical injury on cultured OPCs can be established by applying 10% tensile strain with the FX-4000TTM system.
作者
马祥
祁月红
卫建峰
李燕则
毕竞
吕磊
郭永清
MA Xiang QI Yue-hong WEI Jian-feng LI Yan-ze BI Jing LYU Lei GUO Yong-qing(College of Anaesthesiology,Shanxi Medical University,Taiyuan 030001,China Department of Anaesthesiology,Peopie＇s Hospital Affiliated to Shanxi Medical University in Shanxi Province,Taiyuan 030012,China)
出处
《中国当代医药》
2016年第36期4-7,共4页
China Modern Medicine
基金
山西省自然科学基金项目(2012011042-2)
关键词
少突胶质前体细胞
细胞培养
体外模型
机械拉伸
脊髓损伤
Oligodendrocyte precursor cells
Cell culture
In vitro model
Mechanical strain
Spinal cord injury
