CAJ | 학술논문

本研究目的是从新生SD大鼠海马分离、培养神经干细胞并诱导其向胆碱能神经元方向分化.利用含b-FGF(20 ng/ml)和B27的无血清DMEM/F12培养基培养新生SD大鼠海马分离的具有自我更新和多向分化能力的细胞群,用免疫细胞化学技术检测巢蛋白(nestin),并于分化后分别检查特异性成熟神经细胞、星形胶质细胞、少突胶质细胞的标记抗原β-微管蛋白(Tuj1)、胶质纤维酸性蛋白(GFAP)和半乳糖脑苷脂(Galc)的表达;用鸡胚骨骼肌提取液,诱导神经干细胞向胆碱能神经元方向分化.结果显示:从海马分离的细胞群具有自我更新能力,表达nestin,分化成熟后的细胞表达神经元、星形胶质细胞和少突胶质细胞的特异性抗原;与对照组3.9%相比,鸡胚骨骼肌提取液可以诱导这些细胞中的9.6%分化成为胆碱能神经元.提示分离的细胞具有自我更新能力和多向分化潜能,是中枢神经系统的干细胞;在加有鸡胚骨骼肌提取液的培养基诱导下,能向胆碱能神经元方向分化.
본연구목적시종신생SD대서해마분리、배양신경간세포병유도기향담감능신경원방향분화.이용함b-FGF(20 ng/ml)화B27적무혈청DMEM/F12배양기배양신생SD대서해마분리적구유자아경신화다향분화능력적세포군,용면역세포화학기술검측소단백(nestin),병우분화후분별검사특이성성숙신경세포、성형효질세포、소돌효질세포적표기항원β-미관단백(Tuj1)、효질섬유산성단백(GFAP)화반유당뇌감지(Galc)적표체;용계배골격기제취액,유도신경간세포향담감능신경원방향분화.결과현시:종해마분리적세포군구유자아경신능력,표체nestin,분화성숙후적세포표체신경원、성형효질세포화소돌효질세포적특이성항원;여대조조3.9%상비,계배골격기제취액가이유도저사세포중적9.6%분화성위담감능신경원.제시분리적세포구유자아경신능력화다향분화잠능,시중추신경계통적간세포;재가유계배골격기제취액적배양기유도하,능향담감능신경원방향분화.
The present study aims to isolate neural stem cells from neonatal rat hippocampus and induce them to differentiate into cholinergic neurons. A multipotent cell line derived from the hippocampi of neonatal rats which had the ability to form clones was incubated in serum-free DMEM/F12 medium added with 20ng/ml basic fibroblast growth factor (bFGF) and B27. After differentiation of the neural stem cells, immunocytochemistry was used to detect nestin, the antigen of the cell clone, and β-tubulin (Tuj 1 ), glial fibrillary acidic protein (GFAP) and galactocerebroside (Galc), the markers specific for neurons, astrocytes and oligodendrocytes, respectively. Embryonic chick skeletal muscle extract was used to induce the differentiation of the neural stem cells into cholinergic neurons. The results showed that the cell line isolated from the hippocampi of neonatal rats expressed nestin and had the potential to form clones and differentiate into neurons, astrocytes and oligodendrocytes. Embryonic chick skeletal muscle extract can induce 9.6% of the isolated cell line to differentiate into cholinergic neurons compared with 3.9% in controls. These findings suggested that the cell line, which expressed nestin antigen, was a multipotent cell line capable of self-renewing, and was believed to contain stem cells of the CNS. These neural stem cells can be induced to differentiate into cholinergic neurons by using embryonic chick skeletal muscle extract.