中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
4期
728-730
,共3页
宋平%蔡强%陈谦学%黄书岚%邓钢%易伟%陈治标%郭振涛%吴立权
宋平%蔡彊%陳謙學%黃書嵐%鄧鋼%易偉%陳治標%郭振濤%吳立權
송평%채강%진겸학%황서람%산강%역위%진치표%곽진도%오립권
神经干细胞%细胞培养%海马%大鼠
神經榦細胞%細胞培養%海馬%大鼠
신경간세포%세포배양%해마%대서
Neural stem cells%Cell culture%Hippocampus%Rat
目的 建立新生大鼠海马神经干细胞(NSC)分离培养与鉴定方法.方法 分离新生24 h内SD大鼠海马组织,制成单细胞悬液,采用含表皮生长因子(EGF)和碱性成纤维细胞生长因子(bFGF)的无血清条件进行培养,1周后传代.结果 培养48～ 72 h,细胞聚集悬浮生长,形成典型的神经球,呈神经干细胞巢蛋白(Nestin)阳性表达,并能在体外传代连续形成克隆,原代和传代的NSC加入含10％胎牛血清的DMEM/F12培养基,1～2 d后神经球即开始贴壁分化生长,第7天分别呈神经元特异性烯醇化酶(NSE)阳性和胶质纤维酸性蛋白(GFAP)阳性表达.结论 新生大鼠海马中存在NSC,分离培养时具有自我更新增殖和多分化潜能.
目的 建立新生大鼠海馬神經榦細胞(NSC)分離培養與鑒定方法.方法 分離新生24 h內SD大鼠海馬組織,製成單細胞懸液,採用含錶皮生長因子(EGF)和堿性成纖維細胞生長因子(bFGF)的無血清條件進行培養,1週後傳代.結果 培養48～ 72 h,細胞聚集懸浮生長,形成典型的神經毬,呈神經榦細胞巢蛋白(Nestin)暘性錶達,併能在體外傳代連續形成剋隆,原代和傳代的NSC加入含10％胎牛血清的DMEM/F12培養基,1～2 d後神經毬即開始貼壁分化生長,第7天分彆呈神經元特異性烯醇化酶(NSE)暘性和膠質纖維痠性蛋白(GFAP)暘性錶達.結論 新生大鼠海馬中存在NSC,分離培養時具有自我更新增殖和多分化潛能.
목적 건립신생대서해마신경간세포(NSC)분리배양여감정방법.방법 분리신생24 h내SD대서해마조직,제성단세포현액,채용함표피생장인자(EGF)화감성성섬유세포생장인자(bFGF)적무혈청조건진행배양,1주후전대.결과 배양48～ 72 h,세포취집현부생장,형성전형적신경구,정신경간세포소단백(Nestin)양성표체,병능재체외전대련속형성극륭,원대화전대적NSC가입함10％태우혈청적DMEM/F12배양기,1～2 d후신경구즉개시첩벽분화생장,제7천분별정신경원특이성희순화매(NSE)양성화효질섬유산성단백(GFAP)양성표체.결론 신생대서해마중존재NSC,분리배양시구유자아경신증식화다분화잠능.
Objective To establish the methods for culture and identification of neural stem cells isolated from the hippocampus of neonatal rats.Methods The hippocampal tissues of 24-h newborn SD rats were separated to produce single cell suspension,which was cultured with serum-free medium supplemented with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF),and passaged one week later.Results After primary culture for 48-72 h,cells gathered with suspension growth,and typical neurosphere formed,expressed nestin and could be passaged in vitro.Primary and passage cultured NSCs were cultured in DMEM/F12 medium supplemented with 10％ fetal bovine serum,1-2 days later neurospheres adhered to the wall and differentiated,and at day 7,neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) were expressed.Conclusion The NSCs exist in the hippocampus of newborn rats,and they have potentials of self-renewal and multipotential differentiation during isolation and culture.
