中国实用神经疾病杂志
中國實用神經疾病雜誌
중국실용신경질병잡지
CHINESE JOURNAL OF PRACTICAL NERVOUS DISEASES
2014年
8期
1-5
,共5页
孟磊%周文科%金保哲%惠磊%钟根深%李武雄%王仲伟%黄立勇%张新中
孟磊%週文科%金保哲%惠磊%鐘根深%李武雄%王仲偉%黃立勇%張新中
맹뢰%주문과%금보철%혜뢰%종근심%리무웅%왕중위%황립용%장신중
神经干细胞%细胞培养%成年大鼠%免疫荧光细胞化学
神經榦細胞%細胞培養%成年大鼠%免疫熒光細胞化學
신경간세포%세포배양%성년대서%면역형광세포화학
Neural stem cells%Cell culture%Adult rat%Immunofluorescence cytochemis
目的：改良成年SD大鼠神经干细胞分离、培养及鉴定方法，观察神经干细胞的生长、增殖及分化特点，为后续实验提供细胞。方法从成年SD大鼠分离出完整海马齿状回，采用机械吹打法获得原代细胞，用accutase消化传代，利用cck-8法检测神经干细胞的增殖情况，利用多重免疫荧光细胞化学方法鉴定神经干细胞及其分化细胞。结果机械吹打法可高效获得原代神经干细胞，accutase消化传代更有利于神经干细胞的传代培养，cck-8法简单高效的测定了神经干细胞的增殖，多重免疫荧光创新性的动态展示了神经干细胞经诱导分化后的分化过程。结论改良后的方法可更简单高效的获得和培养出大量细胞，经多重免疫荧光鉴定所分离和培养的细胞是神经干细胞。
目的：改良成年SD大鼠神經榦細胞分離、培養及鑒定方法，觀察神經榦細胞的生長、增殖及分化特點，為後續實驗提供細胞。方法從成年SD大鼠分離齣完整海馬齒狀迴，採用機械吹打法穫得原代細胞，用accutase消化傳代，利用cck-8法檢測神經榦細胞的增殖情況，利用多重免疫熒光細胞化學方法鑒定神經榦細胞及其分化細胞。結果機械吹打法可高效穫得原代神經榦細胞，accutase消化傳代更有利于神經榦細胞的傳代培養，cck-8法簡單高效的測定瞭神經榦細胞的增殖，多重免疫熒光創新性的動態展示瞭神經榦細胞經誘導分化後的分化過程。結論改良後的方法可更簡單高效的穫得和培養齣大量細胞，經多重免疫熒光鑒定所分離和培養的細胞是神經榦細胞。
목적：개량성년SD대서신경간세포분리、배양급감정방법，관찰신경간세포적생장、증식급분화특점，위후속실험제공세포。방법종성년SD대서분리출완정해마치상회，채용궤계취타법획득원대세포，용accutase소화전대，이용cck-8법검측신경간세포적증식정황，이용다중면역형광세포화학방법감정신경간세포급기분화세포。결과궤계취타법가고효획득원대신경간세포，accutase소화전대경유리우신경간세포적전대배양，cck-8법간단고효적측정료신경간세포적증식，다중면역형광창신성적동태전시료신경간세포경유도분화후적분화과정。결론개량후적방법가경간단고효적획득화배양출대량세포，경다중면역형광감정소분리화배양적세포시신경간세포。
Objective To improve method for isolating ,culturing and identifying neural stem cells (NSCs)derived from the hippocampus dentate gyrus in adult SD rat ,observe characteristics of growth ,multiplication and differentiation ,and prepare NSCs for subsequent experiments.Methods The whole hippocampus dentate gyrus was separated from adult SD rat. Primary cells were acquired by making use of mechanical blow. The neural cell pellet were passaged by accutase digestion method ,pro-liferation of NSCs could be determined by cck-8 method. Cultured and differentiated cells were identified with multiple immuno-fluorescence cytochemistry method. Results Primary neural stem cells could be efficiently obtained by mechanical blow ,ac-cutase digestive method was more conducive to the neural stem cells in the process of subculture ,the CCK-8 method was simple and efficient to determine the proliferation of neural stem cells ,multiple immunofluorescence innovatively dynamically displayed the differentiation process of neural stem cells after being induced.Conclusion The improved method is more simple and effi-cient in obtaining and culturing a large number of cells. The isolated and cultured cells are determined to be neural stem cells by multiple immunofluorescence.
