中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
23期
3691-3695
,共5页
夏许可%张伟%余杰锋%章莹
夏許可%張偉%餘傑鋒%章瑩
하허가%장위%여걸봉%장형
干细胞%培养%神经干细胞%干细胞培养%体外诱导分化%免疫组织化学%山羊
榦細胞%培養%神經榦細胞%榦細胞培養%體外誘導分化%免疫組織化學%山羊
간세포%배양%신경간세포%간세포배양%체외유도분화%면역조직화학%산양
stem cells%neural stem cells%cells,cultured%goats
背景:文献报道不同种类的神经调控因子及神经胶质细胞对神经干细胞分化成熟具有不同的作用,但大动物的神经干细胞培养报道则相对较少。目的:探究山羊神经干细胞的培养条件及体外诱导分化结果。方法:取幼羊脑组织分离神经干细胞后于神经干细胞全培养液中悬浮培养,后期行抗-巢蛋白一抗免疫组织化学染色鉴定,同时取坐骨神经分离培养许旺细胞;以血清诱导干细胞体外诱导分化后行抗-S100一抗、抗-胶质纤维酸性蛋白一抗、抗-MAP2一抗染色分别标记许旺细胞及神经胶质细胞,以未添加一抗做对照,计算图像灰度值与对照组进行统计学比较。结果与结论:成功分离培养抗-巢蛋白染色阳性神经球及抗-S100染色阳性许旺细胞,后期神经干细胞体外诱导分化得抗-胶质纤维酸性蛋白、抗-MAP2染色阳性细胞,其免疫组织化学染色灰度值显著高于对照组;分化产物抗-S100染色为阴性,其免疫组织化学染色灰度值显著低于对照组。结果表明山羊神经干细胞可经体外诱导分化为神经元及星形胶质细胞,未发现神经干细胞诱导分化为许旺细胞。
揹景:文獻報道不同種類的神經調控因子及神經膠質細胞對神經榦細胞分化成熟具有不同的作用,但大動物的神經榦細胞培養報道則相對較少。目的:探究山羊神經榦細胞的培養條件及體外誘導分化結果。方法:取幼羊腦組織分離神經榦細胞後于神經榦細胞全培養液中懸浮培養,後期行抗-巢蛋白一抗免疫組織化學染色鑒定,同時取坐骨神經分離培養許旺細胞;以血清誘導榦細胞體外誘導分化後行抗-S100一抗、抗-膠質纖維痠性蛋白一抗、抗-MAP2一抗染色分彆標記許旺細胞及神經膠質細胞,以未添加一抗做對照,計算圖像灰度值與對照組進行統計學比較。結果與結論:成功分離培養抗-巢蛋白染色暘性神經毬及抗-S100染色暘性許旺細胞,後期神經榦細胞體外誘導分化得抗-膠質纖維痠性蛋白、抗-MAP2染色暘性細胞,其免疫組織化學染色灰度值顯著高于對照組;分化產物抗-S100染色為陰性,其免疫組織化學染色灰度值顯著低于對照組。結果錶明山羊神經榦細胞可經體外誘導分化為神經元及星形膠質細胞,未髮現神經榦細胞誘導分化為許旺細胞。
배경:문헌보도불동충류적신경조공인자급신경효질세포대신경간세포분화성숙구유불동적작용,단대동물적신경간세포배양보도칙상대교소。목적:탐구산양신경간세포적배양조건급체외유도분화결과。방법:취유양뇌조직분리신경간세포후우신경간세포전배양액중현부배양,후기행항-소단백일항면역조직화학염색감정,동시취좌골신경분리배양허왕세포;이혈청유도간세포체외유도분화후행항-S100일항、항-효질섬유산성단백일항、항-MAP2일항염색분별표기허왕세포급신경효질세포,이미첨가일항주대조,계산도상회도치여대조조진행통계학비교。결과여결론:성공분리배양항-소단백염색양성신경구급항-S100염색양성허왕세포,후기신경간세포체외유도분화득항-효질섬유산성단백、항-MAP2염색양성세포,기면역조직화학염색회도치현저고우대조조;분화산물항-S100염색위음성,기면역조직화학염색회도치현저저우대조조。결과표명산양신경간세포가경체외유도분화위신경원급성형효질세포,미발현신경간세포유도분화위허왕세포。
BACKGROUND:Different types of nerve regulatory factors and glial cells have been reported to exert different roles in the differentiation and maturation of neural stem cells, but culturing neural stem cells in large animals is relatively rarely reported. OBJECTIVE:To explore the way for culturing goat neural stem cells and to detect the outcome after in vitro differentiation. METHODS:The neural stem cellwas separated and cultured from the newborn goat cerebral cortex and the anti-nestin immunocytochemical staining was performed for cellidentification. At the same time, anti-S100 active Schwann cells were gotten from the sciatic nerve. Then in vitro differentiation was preformed and the outcome was detected by the immunocytochemical stain of anti-glial fibril ary acidic protein, anti-microtubule-associated protein 2 and anti-S100. cells without primary antibodies served as controls. Gray values were calculated and compared. RESULTS AND CONCLUSION:The Schwann cells were cultured successful y, which were active to the anti-nestin immunocytochemical staining and anti-S100 staining. After differentiation, the products were active to anti-glial fibril ary acidic protein and anti-microtubule-associated protein 2 immunocytochemical stain, but Abstract BACKGROUND:Different types of nerve regulatory factors and glial cells have been reported to exert different roles in the differentiation and maturation of neural stem cells, but culturing neural stem cells in large animals is relatively rarely reported. OBJECTIVE:To explore the way for culturing goat neural stem cells and to detect the outcome after in vitro differentiation. METHODS:The neural stem cellwas separated and cultured from the newborn goat cerebral cortex and the anti-nestin immunocytochemical staining was performed for cellidentification. At the same time, anti-S100 active Schwann cells were gotten from the sciatic nerve. Then in vitro differentiation was preformed and the outcome was detected by the immunocytochemical stain of anti-glial fibril ary acidic protein, anti-microtubule-associated protein 2 and anti-S100. cells without primary antibodies served as controls. Gray values were calculated and compared. RESULTS AND CONCLUSION:The Schwann cells were cultured successful y, which were active to the anti-nestin immunocytochemical staining and anti-S100 staining. After differentiation, the products were active to anti-glial fibril ary acidic protein and anti-microtubule-associated protein 2 immunocytochemical stain, but negative to the anti-S100. And significant difference was found in gray values. The goat neural stem cells and Schwann cells were successful y cultured and identified. After the differentiation, the astrocytes and neurons were detected, but the Schwann cells were not found.