中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
9期
1864-1866,封3
,共4页
焦红亮%王晓宁%孙剑瑞%李建斌%关方霞%杨波
焦紅亮%王曉寧%孫劍瑞%李建斌%關方霞%楊波
초홍량%왕효저%손검서%리건빈%관방하%양파
羊膜%间充质干细胞%诱导%混合培养
羊膜%間充質榦細胞%誘導%混閤培養
양막%간충질간세포%유도%혼합배양
Amnion%Mesenchymal stem cells%Induction%Mixed culture
目的 观察羊膜间充质干细胞(HAMs)可诱导分化为神经样细胞混合培养的生长以及经碱性成纤维细胞生长因子(bFGF)诱导HAMs向神经样细胞分化.方法 采用细胞分离和培养技术获取HAMs,分离HAMs后传代培养,加入bFGF诱导分化.当细胞传至第3代时,随机取培养的6份HAMs(半量)为A组,6份HAMs(半量)为B组,将A、B组各剩余的半量HAMs混合为C组.3组细胞密度均为1.0 x 107/L.以活细胞计数和噻唑蓝(MTT)比色法比较3组细胞扩增数量,免疫组织化学法检测HAMs胶质纤维酸性蛋白(GFAP)、神经元特异性烯醇化酶(NSE)、巢蛋白(Nestin)的表达.以活细胞计数和MTT比色法比较3组细胞扩增数量,免疫组织化学检测HAMs神经胶质细胞标志物、神经元特异性烯醇化酶和巢蛋白的表达.结果 活细胞计数结果显示,第18天A组HAMs为(4.80±1.49)×104个,B组HAMs为(5.00±1.75) xl04个,C组HAMs为(8.90±3.46)×104个.MTT比色检测结果均显示,第18天A组HAMs为0.242±3.408,B组HAMs为0.245±5.226,C组HAMs为0.321 ±1.321(P ＜0.05).经bFGF诱导后均表达GFAP 、NSE和Nestin.结论 混合HAMs之间有互相促增殖作用,HAMs具有较强的可塑性,经bFGF诱导的HAMs可表达GFAP、NSE和Nestin.
目的 觀察羊膜間充質榦細胞(HAMs)可誘導分化為神經樣細胞混閤培養的生長以及經堿性成纖維細胞生長因子(bFGF)誘導HAMs嚮神經樣細胞分化.方法 採用細胞分離和培養技術穫取HAMs,分離HAMs後傳代培養,加入bFGF誘導分化.噹細胞傳至第3代時,隨機取培養的6份HAMs(半量)為A組,6份HAMs(半量)為B組,將A、B組各剩餘的半量HAMs混閤為C組.3組細胞密度均為1.0 x 107/L.以活細胞計數和噻唑藍(MTT)比色法比較3組細胞擴增數量,免疫組織化學法檢測HAMs膠質纖維痠性蛋白(GFAP)、神經元特異性烯醇化酶(NSE)、巢蛋白(Nestin)的錶達.以活細胞計數和MTT比色法比較3組細胞擴增數量,免疫組織化學檢測HAMs神經膠質細胞標誌物、神經元特異性烯醇化酶和巢蛋白的錶達.結果 活細胞計數結果顯示,第18天A組HAMs為(4.80±1.49)×104箇,B組HAMs為(5.00±1.75) xl04箇,C組HAMs為(8.90±3.46)×104箇.MTT比色檢測結果均顯示,第18天A組HAMs為0.242±3.408,B組HAMs為0.245±5.226,C組HAMs為0.321 ±1.321(P ＜0.05).經bFGF誘導後均錶達GFAP 、NSE和Nestin.結論 混閤HAMs之間有互相促增殖作用,HAMs具有較彊的可塑性,經bFGF誘導的HAMs可錶達GFAP、NSE和Nestin.
목적 관찰양막간충질간세포(HAMs)가유도분화위신경양세포혼합배양적생장이급경감성성섬유세포생장인자(bFGF)유도HAMs향신경양세포분화.방법 채용세포분리화배양기술획취HAMs,분리HAMs후전대배양,가입bFGF유도분화.당세포전지제3대시,수궤취배양적6빈HAMs(반량)위A조,6빈HAMs(반량)위B조,장A、B조각잉여적반량HAMs혼합위C조.3조세포밀도균위1.0 x 107/L.이활세포계수화새서람(MTT)비색법비교3조세포확증수량,면역조직화학법검측HAMs효질섬유산성단백(GFAP)、신경원특이성희순화매(NSE)、소단백(Nestin)적표체.이활세포계수화MTT비색법비교3조세포확증수량,면역조직화학검측HAMs신경효질세포표지물、신경원특이성희순화매화소단백적표체.결과 활세포계수결과현시,제18천A조HAMs위(4.80±1.49)×104개,B조HAMs위(5.00±1.75) xl04개,C조HAMs위(8.90±3.46)×104개.MTT비색검측결과균현시,제18천A조HAMs위0.242±3.408,B조HAMs위0.245±5.226,C조HAMs위0.321 ±1.321(P ＜0.05).경bFGF유도후균표체GFAP 、NSE화Nestin.결론 혼합HAMs지간유호상촉증식작용,HAMs구유교강적가소성,경bFGF유도적HAMs가표체GFAP、NSE화Nestin.
Objective To observe growth of basic fibroblast growth factor (bFGF)-induced cultures of human amnion-derived mesenchymal stem cells (HAMs) and differentiation into neuronal-like cells.Comparative observation.Methods Amnia from full-term,uterine-incision delivery were donated by 12 healthy women.HAMs were obtained by cell separation and culture techniques,and were passaged and induced by bFGF.From the third passage,a total of 1 ml HAMs,at a density of 1.0 × 107/L,was separately harvested from six samples,which served as group A.A total of 1 ml HAMs,at a density of 1.0 × 107/L,was harvested separately from the remaining six samples,which served as group B.A total of 0.5 ml from the six samples of group A and 0.5 ml from the six samples of group B were combined to form group C.Differences in cell quantity among the three groups were compared by cell quantification and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis.Expressions of a glial cell marker,neuron-specific enolase,and Nestin were detected in the three groups by immunocytochemistry.Results Cell quantification and MTT analysis of live cells,as well as AMSC absorbance,were significantly greater in group C compared with groups A and B at 18 days of culture (P ＜ 0.05),and no significant difference was observed between groups A and B.Glial fibrillary acidic protein,neuron-specific enolase,and Nestin were expressed in all groups following bFGF induction.Conclusion Mixed AMSC cultures promoted proliferation,and bFGF-induced HAMs differentiated into neuronal-like cells.
