中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (1): 59-64.doi: 10.3969/j.issn.2095-4344.0411

• 干细胞移植 stem cell transplantation • 上一篇    下一篇

骨髓间充质干细胞移植对脊髓损伤的修复作用

孙经淞1，周雪颖2，曲淑贤3，李双月2   

  1. 1大连医科大学附属第一医院，辽宁省大连市 116011；大连医科大学，2公共卫生学院，3肿瘤干细胞研究院，辽宁省大连市 116044
  • 修回日期:2017-08-20 出版日期:2018-01-08 发布日期:2018-01-08
  • 通讯作者: 李双月，博士，副教授，大连医科大学公共卫生学院，辽宁省大连市 116044
  • 作者简介:孙经淞，男，1988年生，辽宁省大连市人，汉族，2014年大连医科大学毕业，硕士，主要从事神经毒理相关研究。 并列第一作者：周雪颖，女，1992年生，河北省邢台市人，汉族，2015年大连医科大学毕业，硕士，主要从事干细胞相关研究。

Therapeutic effect of bone marrow mesenchymal stem cell transplantation for spinal cord injury

Sun Jing-song1, Zhou Xue-ying2, Qu Shu-xian3, Li Shuang-yue2   

  1. 1First Affiliated Hospital of Dalian Medical University, Dalian 116011, Liaoning Province, China; 2School of Public Health, Dalian Medical University, Dalian 116044, Liaoning Province, China; 3Institute of Cancer Stem Cells, Dalian Medical University, Dalian 116044, Liaoning Province, China
  • Revised:2017-08-20 Online:2018-01-08 Published:2018-01-08
  • Contact: Li Shuang-yue, M.D., Associate professor, School of Public Health, Dalian Medical University, Dalian 116044, Liaoning Province, China
  • About author:Sun Jing-song, Master, First Affiliated Hospital of Dalian Medical University, Dalian 116011, Liaoning Province, China. Zhou Xue-ying, Master, School of Public Health, Dalian Medical University, Dalian 116044, Liaoning Province, China. Sun Jing-song and Zhou Xue-ying contributed equally to this work.

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摘要：

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文题释义：
全骨髓差速贴壁法的优点：采用全骨髓差速贴壁法分离、纯化和扩增大鼠骨髓间充质干细胞，操作步骤简单，获得骨髓细胞悬液后直接培养，减少了物理筛选或离心对细胞的损伤，极大程度保留了细胞的活性。此外，应用该方法能较完整保留骨髓内微环境，使骨髓间充质干细胞较短时间内贴壁，所得到的骨髓间充质干细胞数量较多，纯度较高。
脊髓损伤：是指外力直接或间接作用于脊髓，引起脊髓功能性或器质性损害，以及继发一系列病理改变进一步加重脊髓损伤，从而出现不可逆性的损伤平面以下感觉、运动、括约肌功能障碍、肌张力异常及病理反射。理想的脊髓损伤模型应具3方面条件：①临床相似性，即制作的脊髓损伤模型与临床相近，尽量接近人类情况；②可调控性，可根据实际需要调整损伤强度，复制出不同损伤程度的脊髓损伤模型；③可操作性与可重复性，即对脊髓损伤模型制作的各关键步骤客观化、定量化，使其可信度高、可实施性强，能反映客观问题。

 

摘要
背景：目前对于脊髓损伤的治疗方法有限，治疗效果不佳，探索新的治疗方法显得至关重要。
目的：观察尾静脉注射骨髓间充质干细胞对大鼠脊髓损伤的治疗效果。
方法：采用全骨髓贴壁法进行体外分离、培养和纯化骨髓间充质干细胞，通过细胞形态学观察、表面标记物检测进行鉴定。将30只SD大鼠随机分为正常组、脊髓损伤组和骨髓间充质干细胞移植组，每组10只。利用动脉瘤夹钳夹闭大鼠第10胸椎的脊髓制备脊髓损伤模型，脊髓损伤模型建立10 min后尾静脉注射0.2 mL骨髓间充质干细胞单细胞悬液(2×106个细胞)。移植后0，10，20，30 d分别进行运动功能评分，并于移植后30 d，分离脊髓组织进行苏木精-伊红染色和透射电镜观察。
结果与结论：①骨髓间充质干细胞在镜下呈长梭形或成纤维细胞样，CD44、CD90阳性表达率均大于90%，CD45的表达不足2%，证实得到的细胞即为骨髓间充质干细胞；②移植后20，30 d，骨髓间充质干细胞移植组大鼠运动功能评分显著高于脊髓损伤组(P < 0.05)；③脊髓损伤组大鼠脊髓组织结构破坏，血管破裂损伤，神经元细胞肿胀变性，并有炎症细胞浸润。骨髓间充质干细胞移植后，脊髓神经元数量明显增多，细胞膜完整，细胞核仁清晰可见；④脊髓损伤组大鼠脊髓轴突肿胀、髓鞘脱失，神经轴索出现变性和坏死。骨髓间充质干细胞移植后，大鼠脊髓轴突结构明显修复，部分脱失的髓鞘恢复，厚度均一；⑤结果表明，骨髓间充质干细胞尾静脉移植对大鼠脊髓损伤有明显的治疗作用，可能与其修复脊髓组织结构、保护轴突和髓鞘结构完整有关。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0002-8030-2062(孙经淞)

关键词: 干细胞, 骨髓间充质干细胞, 脊髓损伤, 干细胞移植, 轴突, 髓鞘

Abstract:

BACKGROUND: At present, treatments for spinal cord injury are limited, with poor outcomes. Therefore, it is of great importance to explore new treatment methods.
OBJECTIVE: To observe the therapeutic effect of bone marrow mesenchymal stem cells (BMSCs) injection via the caudal vein on spinal cord injury in rats.
METHODS: BMSCs were isolated and cultured in vitro by whole bone marrow adherence method. The surface markers were identified by flow cytometry. Thirty Sprague-Dawley rats were randomly divided into control group, spinal cord injury group and BMSCs transplantation group, 10 rats in each group. A rat spinal cord injury model was established by occlusion of the 10th thoracic vertebra using an aneurysm clamp, and 2×106 BMSCs were injected through the caudal vein at 10 minutes after modeling. Basso-Bettle-Bresnahan (BBB) score for motor function recovery was assessed at 0, 10, 20, 30 days after transplantation. The therapeutic effect was evaluated by hematoxylin-eosin staining and electron transmission microscopy at 30 days after implantation.
RESULTS AND CONCLUSION: Under the microscope, fusiformis-shaped or fibroblast-like cells were observed. The expression rate of CD44 and CD90 was more than 90% and the expression of CD45 was less than 2%, by which, the BMSCs were identified. The BBB scores were significantly higher in the BMSCs transplantation group than the spinal cord injury group at 20 and 30 days after transplantation (P < 0.05). Hematoxylin-eosin staining showed that there was spinal cord tissue damage, vascular rupture injury, neuronal cell degeneration and inflammatory cell infiltration in the spinal cord injury group. After BMSCs transplantation, the number of spinal cord neurons was markedly increased with intact cytomembrane and clear nucleolus. Electron microscopic results showed that spinal cord axon swelling, demyelination, nerve axon deformation and necrosis were observed in the spinal cord injury group, while after BMSCs transplantation, the rat spinal cord axon structure was repaired, and partially lost myelin was recovered with uniform thickness. To conclude, BMSCs transplantation via the caudal vein has a significant therapeutic effect on spinal cord injury in rats by repairing the spinal cord structure and protecting the integrity of axon and myelin structures.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cell Transplantation, Spinal Cord Injuries, Tissue Engineering

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