兰州大学机构库 >学院待认领
骨髓间充质干细胞联合定向诱导的神经元
Alternative TitleCo-transplantation of Autologous Neuronal Differentiated and Undifferentiated Mesench
张增山
Thesis Advisor文益民
2011-05-18
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword骨髓间充质干细胞 神经元样细胞 细胞移植 细胞分化 脊髓损伤
Abstract目的:探讨一定条件下骨髓间充质干细胞向神经元样细胞分化的能力,并进一步研究两种细胞联合移植治疗脊髓损伤(spinal cord injury SCI)的效果。 方法:选取3月龄雌性SD大鼠62只,两只处死后提取骨髓间充质干细胞用于细胞培养和分化研究,剩余大鼠采用改良Allen,S法制备T9-11SCI模型,然后随机分为4组(n=15),A组为空白对照组,B组为骨髓间充质干细胞组,C组为神经元样细胞组,D组为联合移植组。诱导后的细胞采用免疫荧光方法检测细胞表面特异性标志,各组细胞用Hoechst33342标记,脊髓损伤模型建立1周后将细胞以局部注射的方法移植到损伤区,细胞移植后的1、2、4、6周对各组动物进行Basso-Beattie-Bresnahan(BBB)评分,并行HE染色、荧光显微镜、免疫荧光法检测细胞在体内的存活和分化。 结果:术后2、4、6周各细胞移植组BBB评分较空白对照组相比显著增加 (P<0.05),D组高于B、C两组 (P<0.05),B、C两组组间比较差异无统计学意义(P>0.05)。HE染色示B、C、D组术后六周的脊髓空洞较A组均减小,以D组最明显;荧光显微镜显示移植的细胞存活于脊髓损伤的区域;免疫荧光染色显示诱导后的细胞与体内存活的细胞表面特异性标志神经元特异性烯醇化酶(neuron specific enolase, NSE)、神经丝蛋白200(neuroflament200, NF-200)表达呈阳性,胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)表达呈弱阳性,其中D组NSE和NF-200阳性细胞数量较多,B、C两组较少。 结论:体内体外一定条件下骨髓间充质干细胞都有向神经元样细胞分化的能力;骨髓间充质干细胞和定向诱导的神经元样细胞都可用于移植治疗脊髓损伤,但以两者联合移植治疗的效果最佳。
Other AbstractObjective To evaluate the capability of bone marrow mesenchymal stem cells(MSCs) differentiation into neuron-like cells, and investigate the effects of a combination of the two cells in treatment of spinal cord injury (SCI) in rats. Methords sixty-two three -months-old female SD rats were selected.Mesenchymal stem cells were taken and their cell differentiation was studied in two rats, the others were made for SCI models at T9-11 levels by using a modified Allen’ s stall,which were randomly divided into 4 groups (n=15,each): control group(A), MSCs transplantation group(B), neurons group (C)and Co-transplantation group(D). Immunofluorescence technology was used to detect the expression of the specific markers of the induced cells. Each group of the cells were labeled by Hoechst33342 and transplanted into the injured spinal cord with local injection first week post-injury.At 1, 2, 4, and 6 weeks of surgery,Basso-Beattie-Bresnahan(BBB)scorces were recorded in order to evaluate the functional recovery of the hindlimbs each group . The presence and differentiation of MSCs in vivo were observed by HE, fluorescence microscopy and immunofluorescence technology. Results At 2, 4, 6 weeks after sugery ,there were significant improvement of BBB scores in groups B, C and D compared with group A(P<0.05),The score of group D was higher than groups B and C(P<0.05).There were no differences between groups B and C(P>0.05). HE staining revealed that the area of spinal cavity in groups B, C and D ,especially in group D,were smaller 6 weeks post-operation compared with group A.The fluorescence microscopy showed that the transplanted cells survived in the injured region; Immunofluorescence of the surface specific marker such as the neuron specific enolase (NSE) and neuroflament 200 (NF-200) were positive in induced cells and transplanted cells survival in vivo .The expression of glial fibrillary acidic protein (GFAP) was just weakly positive. A large number of NSE and NF-200 cells were detected in the lesioned area in group D,making a contrast with group B and C. Conclusion Under certain conditions MSCs could differentiate into neuron-like cells in vitro and in vivo. MSCs and neuron-like cells can be transplanted for the treatment of spinal cord injury, and their co-transplantation may be the best option.
URL查看原文
Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223379
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
张增山. 骨髓间充质干细胞联合定向诱导的神经元[D]. 兰州. 兰州大学,2011.
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