郑州大学学报（医学版）
鄭州大學學報（醫學版）
정주대학학보（의학판）
JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES)
2014年
2期
157-161
,共5页
神经干细胞%Matrigel胶%巢蛋白%β-tubulinⅢ%胶质纤维酸性蛋白%大鼠
神經榦細胞%Matrigel膠%巢蛋白%β-tubulinⅢ%膠質纖維痠性蛋白%大鼠
신경간세포%Matrigel효%소단백%β-tubulinⅢ%효질섬유산성단백%대서
neural stem cell%Matrigel%Nestin%β-tubulinⅢ%glial fibrillary acidic protein%rat
目的：观察Matrigel胶对大鼠海马神经干细胞（ NSCs）增殖和分化的影响。方法：自出生24 h内SD大鼠海马组织提取NSCs，取培养第3代细胞，通过Nestin蛋白免疫细胞化学染色进行鉴定。取第3代细胞分为4组，A组接种于多聚赖氨酸包被24孔板，加入正常培养基；B组接种于多聚赖氨酸包被24孔板，加入含胎牛血清的DMEM/F12培养基；C 组接种于Matrigel 胶内，加入正常培养基；D 组接种于Matrigel 胶内，加入含胎牛血清的DMEM/F12培养基。培养1周后，通过免疫荧光染色方法检测NSCs中巢蛋白（ Nestin）、β-tubulinⅢ、胶质纤维酸性蛋白（GFAP）的表达情况。结果：第3代培养细胞Nestin蛋白阳性，证实为NSCs。 A组NSCs分化缓慢，B、C、D组NSCs分化迅速，C、D两组中NSCs形成立体的网状结构。4组NSCs中Nestin、β-tubulinⅢ、GFAP蛋白免疫荧光染色阳性率差异有统计学意义（ F培养基分别为3918．000、437．023和3395．000， F包被分别为291．234、4833．000和50．081，P均＜0．001），表明B、C、D组NSCs分化显著，且C、D两组中NSCs向神经元分化趋势明显，C组NSCs向神经元分化趋势最强。结论：Matrigel胶具有诱导NSCs向神经元分化的作用。
目的：觀察Matrigel膠對大鼠海馬神經榦細胞（ NSCs）增殖和分化的影響。方法：自齣生24 h內SD大鼠海馬組織提取NSCs，取培養第3代細胞，通過Nestin蛋白免疫細胞化學染色進行鑒定。取第3代細胞分為4組，A組接種于多聚賴氨痠包被24孔闆，加入正常培養基；B組接種于多聚賴氨痠包被24孔闆，加入含胎牛血清的DMEM/F12培養基；C 組接種于Matrigel 膠內，加入正常培養基；D 組接種于Matrigel 膠內，加入含胎牛血清的DMEM/F12培養基。培養1週後，通過免疫熒光染色方法檢測NSCs中巢蛋白（ Nestin）、β-tubulinⅢ、膠質纖維痠性蛋白（GFAP）的錶達情況。結果：第3代培養細胞Nestin蛋白暘性，證實為NSCs。 A組NSCs分化緩慢，B、C、D組NSCs分化迅速，C、D兩組中NSCs形成立體的網狀結構。4組NSCs中Nestin、β-tubulinⅢ、GFAP蛋白免疫熒光染色暘性率差異有統計學意義（ F培養基分彆為3918．000、437．023和3395．000， F包被分彆為291．234、4833．000和50．081，P均＜0．001），錶明B、C、D組NSCs分化顯著，且C、D兩組中NSCs嚮神經元分化趨勢明顯，C組NSCs嚮神經元分化趨勢最彊。結論：Matrigel膠具有誘導NSCs嚮神經元分化的作用。
목적：관찰Matrigel효대대서해마신경간세포（ NSCs）증식화분화적영향。방법：자출생24 h내SD대서해마조직제취NSCs，취배양제3대세포，통과Nestin단백면역세포화학염색진행감정。취제3대세포분위4조，A조접충우다취뢰안산포피24공판，가입정상배양기；B조접충우다취뢰안산포피24공판，가입함태우혈청적DMEM/F12배양기；C 조접충우Matrigel 효내，가입정상배양기；D 조접충우Matrigel 효내，가입함태우혈청적DMEM/F12배양기。배양1주후，통과면역형광염색방법검측NSCs중소단백（ Nestin）、β-tubulinⅢ、효질섬유산성단백（GFAP）적표체정황。결과：제3대배양세포Nestin단백양성，증실위NSCs。 A조NSCs분화완만，B、C、D조NSCs분화신속，C、D량조중NSCs형성입체적망상결구。4조NSCs중Nestin、β-tubulinⅢ、GFAP단백면역형광염색양성솔차이유통계학의의（ F배양기분별위3918．000、437．023화3395．000， F포피분별위291．234、4833．000화50．081，P균＜0．001），표명B、C、D조NSCs분화현저，차C、D량조중NSCs향신경원분화추세명현，C조NSCs향신경원분화추세최강。결론：Matrigel효구유유도NSCs향신경원분화적작용。
Aim:To observe the effect of Matrigel on the proliferation and differentiation of rat hippocampus neural stem cells(NSCs).Methods:NSCs were extracted from newborn SD rats′hippocampus, and the cells after 3 generations were identified through Nestin cytochemical staining .The NSCs were allocated into four groups:group A was inoculated on 24-well plate coated by poly-lysine and added normal culture medium;group B was inoculated on 24-well plate coated by poly-lysine and added DMEM/F12 medium containing fetal bovine serum; group C was inoculated on 24-well plate coated by Matrigel and added normal culture medium;group D was inoculated 24-well plate coated by Matrigel and add-ed DMEM/F12 medium contarning fetal bovine serum .After a week, the expressions of Nestin ,β-tubulinⅢand glial fibril-lary acidic protein(GFAP) by immunofluorescence staining.Results:Nestin protein was positive in the cells after 3 genera-tions, which confirmed that they were NSCs .NSCs in group A differentiated slowly , while group B,C and D differentiated rapidly.Especially, NSCs in group C and D formed three-dimensional network structure .The immunofluorescence staining positive rate of Nestin,β-tubulinⅢ and GFAP among the four groups had statistical significance ( Fmedium =3 918.000, 437.023, and 3 395.000 respectively,Fcoat =291.234,4 833.000, and 50.081 respectively,P<0.001),which showed that NSCs in group B,C and D were significantly differentiated , and the trend of NSCs differentiated into neurons was more obvi-ous in group C,D,especially group C.Conclusion:Matrigel could induce NSCs to differentiate into neurons .
