无血清培养基和有血清培养基对背根神经节生长的差异  

Difference in the culture of dorsal root ganglion cells in serum-free medium and serum medium

作　　者：徐金辉 秦绪祯 张鸿程 马艳霞[2] 齐士斌 赛吉拉夫 Xu Jinhui;Qin Xuzhen;Zhang Hongcheng;Ma Yanxia;Qi Shibin;Saijilafu(First Affiliated Hospital of Soochow University,Suzhou 215000,Jiangsu Province,China;Institute of Orthopaedics,Soochow University,Suzhou 215000,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院,江苏省苏州市215000 [2]苏州大学骨科研究所,江苏省苏州市215000

出　　处：《中国组织工程研究》2021年第13期2043-2048,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81571189,81772353),项目负责人:赛吉拉夫;国家重点研究开发项目(2016YFC1100203);江苏省创新创业项目,项目负责人:赛吉拉夫。

摘　　要：背景:有血清培养基和无血清培养基都曾用来培养背根神经节细胞,但两者之间的差异仍不清楚。目的:探究用无血清培养基是否能完全替代有血清培养基培养背根神经节细胞。方法:取8-10周ICR小鼠背根神经节,并通过胶原酶和胰蛋白酶的处理,之后分为电转有血清组、电转无血清组和非电转有血清组、非电转无血清组,电转组背根神经节细胞加入电穿孔缓冲液和增强型绿色荧光蛋白质粒,进行电穿孔转染。细胞培养3 d,进行Tuj1抗体染色。非电转有血清组和非电转无血清组统计轴突分叉、轴突再生长度、细胞存活数以及轴突再生相关蛋白的表达量;电转有血清组和电转无血清组统计轴突分叉、轴突再生长度、细胞存活数以及电转效率。实验方案经苏州大学附属第一医院实验动物伦理委员会批准。结果与结论:①非电转有血清组和非电转无血清组在轴突分叉、轴突再生长度、细胞存活数以及轴突再生相关蛋白的表达量上均无明显差异(P>0.05);②电转有血清组和电转无血清组在轴突分叉、轴突再生长度、电转效率上均无明显差异(P>0.05);相比于电转有血清组,电转无血清组的细胞存活数明显较低(P<0.05);③电转有血清组和非电转有血清组之间,细胞轴突再生长度无明显变化(P>0.05),非电转有血清组的细胞存活数明显高于电转有血清组(P<0.05);④结果表明,在非电转的条件下,有无血清并不影响背根神经节的体外培养,无血清培养基可以替代有血清培养基;而在电转条件下,无血清培养基会使细胞存活数降低,提示在电转条件下,血清对背根神经节的体外培养有重要作用,因此无血清培养基无法取代有血清培养基。BACKGROUND:Both serum-free and serum media have been used to culture dorsal root ganglion cells,but the difference between the two remains unclear.OBJECTIVE:To explore whether serum-free medium can completely replace serum medium for culture of dorsal root ganglion cells.METHODS:The dorsal root ganglion of ICR mice at 8-10 weeks was taken and treated with collagenase and trypsin.After that,the mice were divided into the electroporation+serum group,electroporation+serum-free group,non-electroporation+serum group and non-electroporation+serum-free group.In the electroporation groups,the dorsal root ganglion cells were transfected with electroporation buffer and enhanced green fluorescent protein particles.Cells were cultured for three days.After Tuj1 antibody staining,in the non-electroporation+serum group and non-electroporation+serum-free group,axon branches,axon regeneration length,number of cell survival and the expression of proteins related to axon regeneration were counted.In the electroporation+serum group and electroporation+serum-free group,axon branches,length of axon regeneration,number of cell survival,and electroporation efficiency were measured.This study was approved by the Laboratory Animal Ethics Committee of the First Affiliated Hospital of Soochow University.RESULTS AND CONCLUSION:(1)In the non-electroporation+serum group and non-electroporation+serum-free group,there was no significant difference in axon branches,axon regeneration length,number of cell survival and the expression of axon regeneration related proteins(P>0.05).(2)In the electroporation+serum group and electroporation+serum-free group,there was no significant difference in axon branches,axon regeneration length and electroporation efficiency(P>0.05).Compared with electroporation+serum group,the number of cell survival of the electroporation+serum-free group was significantly lower(P<0.05).(3)There was no significant change in the length of cell axon regeneration between the electroporation+serum group and the nonelectroporation+se

关 键 词：背根神经节细胞 无血清 培养基 轴突 轴突分叉 细胞存活数 电转 蛋白 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]