摘要
目的探讨骨髓间充质干细胞(mesenchymal stemcells,MSCs)源神经细胞脑内移植对帕金森病(Parkinson s disease,PD)大鼠的治疗作用。方法贴壁培养法分离、培养大鼠骨髓MSCs,脑匀浆上清诱导第3代MSCs向神经细胞分化,采用免疫细胞化学法鉴定诱导分化后细胞的性质,激光共聚焦显微镜检测诱导前后细胞Ca2+浓度变化,6只PD大鼠行纹状体内MSCs源神经细胞移植作为细胞移植组,6只PD大鼠作为对照组。细胞移植术后4周检测PD大鼠的行为变化,观察移植细胞在脑内的分布情况。结果倒置显微镜下可见MSCs呈纺锤形和多角形,有1～2个核仁,MSCs经脑匀浆上清诱导后其胞体折光性增强,发出数个细长突起,互相交织成网,有的似轴突。诱导后细胞表达神经元特异性标志物神经元特异性烯醇化酶(NSE)和神经丝蛋白(NF),胞质Ca2+荧光强度显著增强,可推测诱导后的细胞为MSCs源神经细胞,将BrdU标记的MSCs源神经细胞移植到PD大鼠纹状体治疗4周后,可见细胞散在分布于注射侧脑组织,有少量细胞可迁移到对侧脑组织,PD大鼠的旋转行为得到显著改善。结论MSCs源神经细胞移植治疗帕金森病大鼠可使其旋转行为得到改善。
Objective To explore the effects of intracerebal implantation of bone marrow mesenchymal stem cell（MSCs）-derived neurons on Parkinson＇s disease（PD） in 6-OHDA-induced rat model of Parkinson＇s disease.Methods The rat MSCs from bone marrow were cultured by adhering to the culture plastic and identified by immunocytochemistry.Neurons from the MSCs were induced by brain homogenate supernatant treatment in the third passage of culture.The concentrations of intracellular Ca2＋ before and after induction were detected by laser scanning confocal microscopy.Six PD model rats with implanted MSC-derived neurons were included in the transplantation group,and the other six PD model rats as controls.Four weeks later the pathological changes of PD rats were examined and the survival of the transplanted cells in the corpus striatum was assessed.Results Under the inverted microscope the MSCs were spindle and polygonal in shape with a central nucleus containing 1 or 2 nucleoli.The MSCs became round and bright after induction with brain homogenate supernatant for 24 hours.There were several processes projecting from the cell body,and some processes appeared like axons or dendrites.As the cells expressed neuron-specific enolase（NSE） and neurofilament（NF） protein,specific markers of mature neurons,it was considered that neurons were induced from the bone marrow-derived MSCs.The cytoplasmic Ca2＋ fluorescence intensity in the induced cells were significantly stronger than that in the non-induced cells.At 4 weeks after BrdU labeled MSCs-derived neurons were implanted into the striatum of PD rats,scattered BrdU-positive cells were observed in the brain tissue at the implantation side,and a few cells immigrated into the cerebral tissue at the opposite side.The frequency of rotation of PD rats in transplantation group was decreased.Conclusion Implanted MSCs can survive in the brain of PD rat and the rotational behavior of PD rats can be improved.
出处
《中国实验动物学报》
CAS
CSCD
2009年第2期130-134,I0004,共6页
Acta Laboratorium Animalis Scientia Sinica
基金
山东省自然科学基金(Y2003C01)
潍坊医学院博士启动基金
关键词
间充质干细胞
帕金森病
细胞移植
骨髓
大鼠
Mesenchymal stem cells
Parkinson＇s disease
Cell implantation
Bone marrow
Rats
