丁羟茴醚诱导幼儿骨髓间质干细胞分化为神经细胞  

The differentiation of neural cells with child bone marrow stromal cells in vitro

作　　者：师红[1] 季泽娟[2] 刘福云[2] 

机构地区：[1]郑州大学第三附属医院药剂科,河南郑州450052 [2]郑州大学第三附属医院小儿骨科,河南郑州450052

出　　处：《河南医学研究》2010年第3期287-290,共4页Henan Medical Research

摘　　要：目的:探讨幼儿骨髓间质干细胞诱导分化为神经干细胞及神经细胞的方法及生物学特征,为临床利用骨髓间质干细胞或/和造血干细胞治疗小儿脊髓栓系综合征及其它神经系统疾病奠定基础。方法:取健康幼儿骨髓分离纯化骨髓间质干细胞,在α-MEM培养基(含20%胎牛血清,2 mmol/L谷氨酰胺,100 U/ml盘尼西林,100 mg/ml链霉素,25 ng/ml两性霉素B)中进行无分化培养,将传代细胞用丁羟茴醚(ButylatedHydroxyanisole,BHA)诱导,诱导剂是无血清DMEM培养基:含2%二甲基亚砜(Dimethyl sulfoxide,DMSO)、150μMBHA。结果:骨髓间质干细胞经过7 d体外诱导,大约有55%的细胞发生形态学变化,最初是胞体变圆,随后延展出长长的神经丝并相互交织成网,免疫组化结果表明这些细胞beta-微管蛋白Ⅲ(β-tublinⅢ)和神经胶质原纤维酸性蛋白(glial fibrillary acidic protein,GFAP)表达阳性。结论:儿童骨髓间质干细胞在合适的诱导剂作用下体外可定向分化成神经元和神经胶质细胞。为小儿脊髓栓系综合征及其它神经系统疾病自体骨髓移植实验治疗奠定了基础。Objective： This study was designed to investigate the feasibility of differentiation of children marrow stromal stem cells（MSCs） into neural stem cells（NSCs） and neural cells in vitro induced by butylated hydroxyanisole（BHA）.Methods： The MSCs donated voluntarily by a fourteen girl were originally cultured in α-MEM supplemented with 20% FBS,2 mmol/L L-glutamine,100 U/ml penicillin,100 mg/ml streptomycin,and 25 ng/ml amphotericin B.The cells that were expanded as undifferentiated cells in culture for several passages were used to be introduced into neural stem cells（NSCs）and neural cells.The induction media was composed of DMEM,2%DMSO and 150μM BHA.Results： After induction by BHA for 7d,about 35% of the MSCs extended long processes terminating in typical growth cones and filopodia and processes formed extensive networks.The induced cells expressed both beta-tublin Ⅲ and Glial fibrillary acidic protein.In addition,the differentiating cells expressed nestin,characteristic of neuronal precursor stem cells after 30 min,but the trait was undetectable after 6 days.In contrast,expression of trkB,the nerve growth factor receptor,persisted from 5.5 hr to 6 days.Conclusion： It is capable that the MSCs of the Child differentiate into neurons and glial cells with an optimal differentiation protocol.It also is possible to provide abundant the cells which differentiated from the stem cell for treatment of tethered cord syndrome and other neurological diseases in children.

关 键 词：骨髓间质细胞 神经元诱导分化 干细胞 丁羟茴醚 

分 类 号：R72[医药卫生—儿科]