DOI:10．3969/j．issn．1672-7770．2023．02．013·论著·一种小鼠背根神经节原代神经元细胞分离与培养的优化方法张皞晟，王培民【摘要】目的探索一种可以大量快速培养活力好且卫星胶质细胞污染少的原代神经元细胞的方法。方法混合酶溶液消化体式显微镜下提取的背根神经节(DRG)，随后接种于纤维连接蛋白提前包被的培养皿中，8h后更换含5-氟尿嘧啶的维持培养基，抑制非神经元细胞增殖。结果分离培养的感觉神经元细胞在培养过程中可维持15d左右，部分细胞可维持更长时间。原代细胞于第3天即可观察到交错的轴突网络，接种细胞密度大，卫星胶质细胞污染情况少，细胞在7d之内可投入使用，操作者经短时间训练后即可快速掌握方法。结论新方法在传统方法的基础上进行了改良，可获取较多数量细胞活性好、纯度高的原代神经元细胞，相较传统方法具有可操作性高，重复率高的优势。【关键词】背根神经节;神经元细胞;细胞培养;TRPA1【中图分类号】Q813．1+1;R651【文献标志码】A【文章编号】1672-7770(2023)02-0189-06AnoptimizedmethodforisolationandcultureofprimaryneuronsfrommousedorsalrootganglionZHANGHao-sheng，WANGPei-min．InstituteofModernBiology，NanjingUniversity，Nanjing210022，ChinaCorrespondingauthor:WANGPei-minAbstract:ObjectiveToexploreamethodthatcanrapidlycultivatealargenumberofprimaryneuronswithgoodvitalityandlesspollutionofsatelliteglialcells．MethodsDorsalrootganglion(DRG)extractedundermicroscopewasdigestedwithmixedenzymesolution，andtheninoculatedintofibronectincoatedculturedish．After8hours，themaintenancemediumcontaining5-fluorouracilwaschangedtoinhibittheproliferationofnon-neuronalcells．ResultsTheisolatedandculturedsensoryneuroncellscouldlastforabout15days，andsomeofthemcouldlastlonger．Onthethirdday，theprimarycellscouldobservethecrisscrossaxonnetwork．Thedensityoftheinoculatedcellswashigh，andthepollutionofsatelliteglialcellswasless．Thecellscouldbeputintousewithin7days．Theoperatorcouldquicklymasterthemethodafterashortperiodoftraining．ConclusionsThenewmethodisimprovedonthebasisofthetraditionalmethod，whichcanobtainalargenumberofprimaryneuronswithgoodcellactivityandhighpurity．Comparedwiththetraditionalmethod，thenewmethodhasthead...
