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骨髓间充质干细胞和嗅黏膜神经干细胞共移植治疗大鼠脊髓损伤
杨成伟,贾彬莉,甄平,杨新乐,王明,徐雅洁
0
(联勤保障部队第940医院脊柱外科;原兰州军区联勤部通信站卫生所;兰州大学第二医院;河南省荣军医院)
摘要:
【摘要】目的 探讨骨髓间充质干细胞(BMSCs)和嗅黏膜神经干细胞(OMNSCs)共移植对大鼠脊髓损伤后运动功能的影响。方法 分别提取培养并鉴定大鼠BMSCs和OMNSCs。将60只雄性SPF级SD大鼠随机分为假手术组(仅做椎板切除,不损伤脊髓),对照组(在脊髓损伤区域注射5 μL生理盐水),BMSCs移植组(在脊髓损伤区域注射5 μL 5×104个细胞BMSCs单细胞悬液),OMNSCs移植组(在脊髓损伤区域注射5 μL 5×104个细胞 OMNSCs单细胞悬液,BMSCs和OMNSCs共移植组(在脊髓损伤区域注射5μL 1〖DK〗∶1 5×104个细胞BMSCs和OMNSCs细胞混悬液)。细胞移植后第1、3、7、14、21、28天分别对大鼠进行BBB评分,28天后处死大鼠,分离脊髓组织,进行免疫荧光染色观察细胞迁移情况。结果 造模后,大鼠双后肢瘫痪,BBB评分0分。细胞移植后14天起,与对照组相比,干细胞移植组运动功能有显著改善,BBB评分比较差异具有统计学意义(P<005)。与BMSCs移植组和OMNSCs移植组相比,移植后21、28天,共移植组BBB评分显著增加(P<005)。免疫荧光观察结果表明,脊髓损伤28天后,损伤区脊髓内可见空泡形成,两种干细胞聚集在空泡周围,BMSCs在损伤区分布较为集中,而OMNSCs相对散在分布,细胞较为伸展。结论 干细胞移植可以一定程度上恢复脊髓损伤大鼠的运动功能,BMSCs和OMNSCs两种干细胞共移植效果优于单细胞移植。
关键词:  骨髓间充质干细胞  嗅黏膜神经干细胞  移植  脊髓损伤  BBB评分
DOI:
基金项目:国家自然科学基金(81601905);中国博士后科学基金(2017M623433);全军医学科技青年培育项目(15QNP012)
Bone marrow mesenchymal stem cells and olfactory mucous neural stem cells co transplanted to treat spinal cord injury in rats
YANG Chengwei,JIA Binli,ZHEN Ping,YANG Xinle,WANG Ming,XU Yajie
(Spine Surgery, The 940th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army;Former Communication Station Health Office of Joint Service Department of Lanzhou Military Region;The Second Hospital of Lanzhou University;Henan Rongjun Hospital)
Abstract:
【Abstract】Objective To investigate the effect of cotransplantation of bone marrow mesenchymal mtem cells (BMSCs) and olfactory mucous neural stem cells (OMNSCs) on motor function after spinal cord injury in rats. Methods Rat BMSCs and OMNSCs were extracted and cultured. 60 male specific pathogen free (SPF) grade sprague dawley (SD) rats were randomly divided into (1) control group which only underwent laminectomy without damaging the spinal cord; (2) sham group which 5μL of normal saline was injected into the injured area; (3) BMSCs transplantation group which 5μL BMSCs single cell suspension (5×104 cells) was injected into the injured area; (4) OMNSCs transplantation group which 5μL OMNSCs single cell suspension (5×104 cells) was injected into the injured area; (5) BMSCs and OMNSCs cotransplantation group which 5μL BMSCs and OMNSCs (1:1) cell mixture (5×104 cells) was injected into the injured area. Bassobeattlebresnahan (BBB) scores were evaluated on 1, 3, 7, 14 and 28 days after cell transplantation, respectively. After 28 days, the rats were killed and spinal cord tissue was isolated and the immunofluorescence staining was used to observe the cell migration. Results After modeling, the rats' hind limbs were paralyzed, and the BBB score was 0 points. At 14 days after transplantation, the motor function of the stem cell transplantation group was significantly improved compared with the simple spinal cord injured, and the BBB score was statistically significant (P<005). Compared with the BMSCs transplantation group and the OMNSCs transplantation group, the BBB score of the cotransplant group was significantly increased at 21, 28 days after transplantation (P<005). The results of immunofluorescence observation showed that 28 days after spinal cord injury, vacuoles were found in the spinal cord of the injured area, and two kinds of stem cells gathered around the vacuoles. The distribution of BMSCs in the damaged area was more concentrated, while the OMNSCs was relatively scattered, and the OMNSCs were more extended. Conclusion Stem cell transplantation can restore motor function of spinal cord injury rats to a certain extent. The cotransplantation effects of BMSCs and OMNSCs stem cells are better than single cell transplantation.
Key words:  Bone marrow mesenchymal stem cells  Olfactory mucous neural stem cells  Transplantation  Spinal cord injury  BBB score

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