医药前沿
醫藥前沿
의약전연
YIAYAO QIANYAN
2015年
21期
34-35,36
,共3页
背根神经节%动物实验%细胞培养%纯化
揹根神經節%動物實驗%細胞培養%純化
배근신경절%동물실험%세포배양%순화
Dorsal root ganglion%Animal experimentation%Cell Culture%Purification
目的：建立一种简单、稳定、高效的新生大鼠背根神经节神经元原代培养方法。方法：摘取新生24h SD大鼠背根神经节，采用0.25%胰酶和0.1%Ⅳ型胶原酶消化，制成单细胞悬液，接种于Neurobasal/B27无血清培养液中。将培养3d的DRGn于倒置相差显微镜下进行形态学观察，扫描电镜行细胞形态学检测，应用β-tubulinⅢ进行免疫细胞化学染色，鉴定细胞纯度。结果：体外培养的背根神经节神经元生长状态良好，纯度可达到(92±6)%。结论：本实验方法简单、稳定、高效，可以获得高纯度的背根神经节神经元。
目的：建立一種簡單、穩定、高效的新生大鼠揹根神經節神經元原代培養方法。方法：摘取新生24h SD大鼠揹根神經節，採用0.25%胰酶和0.1%Ⅳ型膠原酶消化，製成單細胞懸液，接種于Neurobasal/B27無血清培養液中。將培養3d的DRGn于倒置相差顯微鏡下進行形態學觀察，掃描電鏡行細胞形態學檢測，應用β-tubulinⅢ進行免疫細胞化學染色，鑒定細胞純度。結果：體外培養的揹根神經節神經元生長狀態良好，純度可達到(92±6)%。結論：本實驗方法簡單、穩定、高效，可以穫得高純度的揹根神經節神經元。
목적：건립일충간단、은정、고효적신생대서배근신경절신경원원대배양방법。방법：적취신생24h SD대서배근신경절，채용0.25%이매화0.1%Ⅳ형효원매소화，제성단세포현액，접충우Neurobasal/B27무혈청배양액중。장배양3d적DRGn우도치상차현미경하진행형태학관찰，소묘전경행세포형태학검측，응용β-tubulinⅢ진행면역세포화학염색，감정세포순도。결과：체외배양적배근신경절신경원생장상태량호，순도가체도(92±6)%。결론：본실험방법간단、은정、고효，가이획득고순도적배근신경절신경원。
Objective To establish an simple, efficient, reliable method for the purification culture system of dorsal root ganglion neurons derived from new born rats.Methods Dorsal root ganglions harvested from new born SD rats were digested with the mixture of trypsin and collegoneaseⅣ, then turned into single cell suspension and plated in neuralbasal media. The purified rate was evaluated according to cell count and β-tubulinⅢ immunocytochemistry stain.Results Cultured dorsal root ganglion cells could survive healthily. The purification rate of neurons was(92±6)%.Conclusion The method, which is used for culture and purification of DRGn, is a simple, efficient and reliable way. Using it could obtain highly purify neurons.
