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纸质出版:2013
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赵文勇,万立华,粟永萍,冉新泽.间充质祖细胞源性的神经元样细胞肌内移植后自身特性的维持[J].陆军军医大学学报(原第三军医大学学报),2013,35(08):744-749.
Zhao Wenyong,Wan Lihua,Su Yongping,Ran Xinze.Characteristic maintenance of mesenchymal progenitor cells-derived neuron-like cells after being transplanted into muscle[J].J Amry Med Univ (J Third Mil Med Univ),2013,35(08):744-749.
赵文勇,万立华,粟永萍,冉新泽.间充质祖细胞源性的神经元样细胞肌内移植后自身特性的维持[J].陆军军医大学学报(原第三军医大学学报),2013,35(08):744-749. DOI:
Zhao Wenyong,Wan Lihua,Su Yongping,Ran Xinze.Characteristic maintenance of mesenchymal progenitor cells-derived neuron-like cells after being transplanted into muscle[J].J Amry Med Univ (J Third Mil Med Univ),2013,35(08):744-749. DOI:
目的 探讨间充质祖细胞(mesenchymal progenitor cells,MPCs)源性的神经元样细胞肌内移植后能否维持其自身特性。 方法 取3周龄健康GFP转基因C57小鼠后肢长骨进行MPC培养及鉴定,选取生长良好的第3代MPC,采用神经元原代培养上清液进行神经元样细胞诱导分化后收集细胞,并制备成5×10
5
/μL细胞悬液备用。选取12周龄健康C57小鼠24只,按随机数字表法分为损伤+移植细胞组、损伤+生理盐水组、假手术+移植细胞组、假手术+生理盐水组。损伤+移植细胞组及假手术+移植细胞组:将制备的5 μL MPC悬液散点注入小鼠三头肌内;损伤+生理盐水组和假手术+生理盐水组:同法注入等量生理盐水。术后4周取材,荧光显微镜下观察肌内移植细胞存活情况,电镜观察肌肉超微结构的变化,免疫荧光染色检测神经元特异性标志物NSE、NeuN及神经胶质特异性标志物GFAP的表达情况。 结果 肌内移植细胞生长良好且均匀分布于肌细胞间隙,并抑制了肌细胞核、线粒体、内质网的退变及肌肉纤维化的发生。损伤+移植细胞组定植存活的移植细胞阳性表达:NSE(58.35±1.37)%、NeuN(60.22±0.16)%及GFAP(13.32±1.65)%,损伤+生理盐水组、假手术+移植细胞组、假手术+生理盐水组均无阳性细胞表达。 结论 MPC具有良好的生物学活性
经体外诱导分化为神经元及神经胶质样细胞后移植于失神经支配肌内可定植存活,并能够维持神经元及神经胶质样细胞的特性,同时有效抑制了肌细胞核、线粒体、内质网的退变及肌肉纤维化的发生。
Objective To study the characteristic maintenance of mesenchymal progenitor cells (MPCs)-derived neuron-like cells after being transplanted into denervated muscle. Methods MPCs were separated from bones of hind limbs of GFP transgenic C57 mice mechanically
and were cultured and induced to differentiate in vitro. After being induced
directionally by the supernatants from primary culture of neuron
MPCs at passage3 in good growth were collected and prepared into cell suspension (5×10
5
/μL). Twenty-four 12-week-old C57 mice were randomized into four groups (n=6). In the injury+transplantation and the sham operation + transplantation groups
5 μL of MPCs suspension was injected by points into triceps muscle of mice. In the injury + sodium chloride and the sham operation+sodium chloride group
5 μL of sodium chloride was injected by points into triceps muscle of mice. At 4 weeks after operation
the survival of the transplanted MPC cells were observed by fluorescent microscopy
the ultrastructure was observed by electron microscopy
and the expression of neuronal specific markers of neuron-specific enolase (NSE) and neuronal nuclei protein (NeuN) and neuroglial specific marker of glial fibrillary acidic protein (GFAP) were detected by immunofluorescence staining. Results The transplanted cells grew well and distributed uniformly in the gap of muscle cells. The degeneration of myocyte nucleus
mitochondria and sarcoplasmic reticulum and the musculus fibration were inhibited. NSE [(58.35±1.37)%
]
NeuN [(60.22±0.16)%
]
and GFAP [(13.32±1.65)%
]
were positively expressed in the transplanted cells of the injury + transplantation group rather than in those of other groups. Conclusion MPCs have good biological activity. Neuron-like cells and neuroglia-like cells differentiated from MPCs in vitro can maintain their characteristics and inhibit the degeneration of myocyte nucleus
mitochondria and sarcoplasmic reticulum and musculus fibration after being transplanted into the denervated muscle.
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