[1]马娇,张雪梅,孙杨,等.PKH67示踪骨髓间充质干细胞迁移至损伤脊髓的实验研究[J].卒中与神经疾病杂志,2016,23(06):432-437.[doi:10.3969/j.issn.1007-0478.2016.06.012]
　Ma Jiao,Zhang Xuemei,Sun Yang,et al.Study on the utilization of PKH67 in the transplantation of bone marrow mesenchymal stem cells to injured spinal cord[J].Stroke and Nervous Diseases,2016,23(06):432-437.[doi:10.3969/j.issn.1007-0478.2016.06.012]
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PKH67示踪骨髓间充质干细胞迁移至损伤脊髓的实验研究()
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《卒中与神经疾病》杂志[ISSN:1007-0478/CN:42-1402/R]

卷:
第23卷
期数:
2016年06期
页码:
432-437
栏目:
论 著
出版日期:
2016-12-30

文章信息/Info

Title:
Study on the utilization of PKH67 in the transplantation of bone marrow mesenchymal stem cells to injured spinal cord
文章编号:
1007-0478(2016)06-0432-06
作者:
马娇; 张雪梅; 孙杨; 俞冰倩
150086 哈尔滨医科大学附属第二医院[马娇 张雪梅(通信作者)孙杨 俞冰倩]
Author(s):
Ma Jiao; Zhang Xuemei; Sun Yang; et al.
Department of Neurology,Second Affiliated Hospital of Harbin Medical University,Harbin 150086
关键词:
PKH67 骨髓间充质干细胞 脊髓损伤 增值
Keywords:
PKH67 Bone marrow mesenchymal stem cells Spinal cord injury Proliferation
分类号:
R744
DOI:
10.3969/j.issn.1007-0478.2016.06.012
文献标志码:
A
摘要:
目的 探讨SCI后体外移植PKH67标记的BMSCs迁移至脊髓损伤处并进行增值和分化的动员情况。方法 用梯度离心法分离和培养出SD 大鼠第3代BMSCs,用绿色荧光染料PKH67标记; 采用钳夹法制备脊髓损伤(SCI)模型,分为实验组(n=15)、对照组(n=16)、假手术组(n=16); SCI术后对脊髓损伤组织进行HE染色,实验组和假手术组于术后尾静脉移植含有1×107个BMSCs的0.5 mL生理盐水,对照组注射等量生理盐水; 分别于术后1、7、14、21 d观察大鼠后肢运动功能恢复情况,并做BBB分; 术后21 d后取脊髓组织,行免疫荧光染色,观察BMSCs的迁移,增值和分化情况。结果(1)镜下可见损伤脊髓形成的空洞、坏死及炎性细胞的增多;(2)共聚焦荧光显微镜观察显示术后21 d实验组脊髓损伤部位可见移植的BMSCs, 部分BMSCs呈GFAP和Nestin阳性表达; 假手术组无 PKH67标记的BMSCs; 实验组GFAP和Nestin阳性细胞数较对照组和假手术组明显增加(P<0.05),对照组较假手术组增加不明显(P>0.05);(3)实验组和对照组BBB评分均有增加,但实验组BBB评分显著高于对照组(P<0.05)。结论 PKH67示踪的BMSCs可迁移至损伤脊髓部位,进行增值并分化为神经元样细胞,促进损伤脊髓的神经功能恢复。
Abstract:
ObjectiveTo investigate the utilization of PKH67 in the transplantation of bone marrow mesenchymal stem cells to injured spinal cord and the migration,proliferation and differentiation of the BMSCs.Methods Density gradient centrifugation method was used for isolating the rat BMSCs and the 3rd passage of BMSCs were labeled with green fluorescent dye PKH67.SCI model was generated by using aneurysmclip to clamp the the spinal cord of the rat.There were three groups: BMSCs group,sham operation group and control group.After SCI,the lesion of spinal cord was taken for HE,1.0×107cells labeled with PKH67 in 0.5 mL 0.9% sterile saline solution were transfused intravenously in BMSCs group and sham-operation group,the same volume of sterile saline solution were transfused in control group.The BBB scale was performed at 1 day,7 days,14 days,21 days after surgery for evaluating hind limbs movement.The migration and differentiation of BMSCs were determined by using triple immunofluorescent staining.Results(1)Syringomyelia after injury,necrosis and increased inflammatory cells could be seen under microscopy.(2)Immunofluoresence analysis were performed at 21 days post-transplantation and the transplanted BMSCs were centralized in the epicenter of injured spinal cord,part of BMSCs expressed by GFAP and Nestin.There were no PKH67+ cells in sham operation group.The GFAP+ and Nestin+ cells in BMSCs group were significantly higher than other two groups(P<0.05),while there was no obviously difference between sham operation group and control group(P>0.05).(3)The BBB scores of BMSCs group and control group both improved,but the rats in BMSCs group improved rapidly than control group(P<0.05).Conclusion The bone marrow mesenchymal stem cells labeled with PKH67 could migrate to the lesion of spinal cord,proliferate and differentiate into neurocyte-like cells,which could promote the recovery of spinal cord's function.

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备注/Memo

备注/Memo:
基金项目:于维汉院士杰出青年培养基金项目
更新日期/Last Update: 2016-12-20