济宁医学院学报
濟寧醫學院學報
제저의학원학보
JOURNAL OF JINING MEDICAL COLLEGE
2014年
5期
313-315,319
,共4页
神经干细胞%神经元%分化%促红细胞生成素
神經榦細胞%神經元%分化%促紅細胞生成素
신경간세포%신경원%분화%촉홍세포생성소
Neural stem cells%Culture,neuron%Differentiation%Erythropoietin
目的：探讨促红细胞生成素（erythropoietin ，EPO）促进神经干细胞（neural stem cells ，NSCs）分化的形态学表现，为细胞移植治疗脑部疾病和脑损伤提供实验依据。方法取大鼠14 d胚胎脑皮质先增殖培养，后贴壁分化培养。适时镜下观察，免疫细胞化学染色，用nestin鉴定NSCs ，GFAP鉴定神经胶质细胞、MAP2鉴定神经元。选取第3代 NSCs ，向培养基中分别添加不同剂量的 EPO ，浓度分别为0．5u/ml ，5u/ml ，50u/ml ，500u/ml ，并设不添加EPO的对照组，MAP2免疫荧光共聚焦显微镜观察NSCs向神经元方向的分化。结果1）鼠胚脑皮质在无血清悬浮培养形成大量神经球，并可传代，球内细胞均表达 nestin。分化培养，可检测出MAP2或GFAP阳性细胞。2）EPO≥5u/ml各组分化培养，表达MAP2阳性细胞显著升高（P＜0．05）。结论大鼠胚胎脑皮质体外培养可获得NSCs ；适当浓度EPO可提高NSCs向神经元的分化。
目的：探討促紅細胞生成素（erythropoietin ，EPO）促進神經榦細胞（neural stem cells ，NSCs）分化的形態學錶現，為細胞移植治療腦部疾病和腦損傷提供實驗依據。方法取大鼠14 d胚胎腦皮質先增殖培養，後貼壁分化培養。適時鏡下觀察，免疫細胞化學染色，用nestin鑒定NSCs ，GFAP鑒定神經膠質細胞、MAP2鑒定神經元。選取第3代 NSCs ，嚮培養基中分彆添加不同劑量的 EPO ，濃度分彆為0．5u/ml ，5u/ml ，50u/ml ，500u/ml ，併設不添加EPO的對照組，MAP2免疫熒光共聚焦顯微鏡觀察NSCs嚮神經元方嚮的分化。結果1）鼠胚腦皮質在無血清懸浮培養形成大量神經毬，併可傳代，毬內細胞均錶達 nestin。分化培養，可檢測齣MAP2或GFAP暘性細胞。2）EPO≥5u/ml各組分化培養，錶達MAP2暘性細胞顯著升高（P＜0．05）。結論大鼠胚胎腦皮質體外培養可穫得NSCs ；適噹濃度EPO可提高NSCs嚮神經元的分化。
목적：탐토촉홍세포생성소（erythropoietin ，EPO）촉진신경간세포（neural stem cells ，NSCs）분화적형태학표현，위세포이식치료뇌부질병화뇌손상제공실험의거。방법취대서14 d배태뇌피질선증식배양，후첩벽분화배양。괄시경하관찰，면역세포화학염색，용nestin감정NSCs ，GFAP감정신경효질세포、MAP2감정신경원。선취제3대 NSCs ，향배양기중분별첨가불동제량적 EPO ，농도분별위0．5u/ml ，5u/ml ，50u/ml ，500u/ml ，병설불첨가EPO적대조조，MAP2면역형광공취초현미경관찰NSCs향신경원방향적분화。결과1）서배뇌피질재무혈청현부배양형성대량신경구，병가전대，구내세포균표체 nestin。분화배양，가검측출MAP2혹GFAP양성세포。2）EPO≥5u/ml각조분화배양，표체MAP2양성세포현저승고（P＜0．05）。결론대서배태뇌피질체외배양가획득NSCs ；괄당농도EPO가제고NSCs향신경원적분화。
Objective Methods Results Conclusion To investigate the promotion of EPO on the differenti-ation of the neural stem cells (NSCs) in vitro from cerebral cortex of pregnant SD rats .The brain cortex were iso-lated and cultured in serum-free suspension ,and then in differentiating suspension .Nestin was used to detect the NSCs .Glia fibrillary acid protein (GFAP) and microtubule-associated protein 2 (MAP2 ) were used to detect the dif-ferentiation of NSCs by immunocytochemistry .After obtained the third passage (P3 ) of NSCs ,EPO of different concentration (0.5 ,5 ,50 ,500u/ml) was added to the medium .And there was a control group without EPO .MAP2 was used to detect the neuron after the differentiation by immunocytochemistry .The results showed that the sepa-rated cells from embryonic brain cortex formed neurospheres ,in which nestin-positive cells were detected .GFAP and MAP2-positive cells were detected after differentiation culture .MAP-2-positive cells of the EPO supplementa-tion groups (>5U/ml) showed an significant increase in the immunochemistry dying test compared with the con-trol group .The results indicate that NSCs can be obtained from the separated cerebral cortex of embryonic rats .Ap-propriate concentration of EPO can increase the rate of the neuron after the differentiation the NSCs in vitro .
