天津医药

天津医药 ›› 2015, Vol. 43 ›› Issue (7): 749-752.doi: 10.11958/j.issn.0253-9896.2015.07.013

• 实验研究 • 上一篇    下一篇

不同时期移植人脐血 CD34+细胞对大鼠脊髓损伤修复的对比研究

唐亮，冯世庆，高瑞霄   

  1. 1天津医科大学总医院 （邮编 300052）； 2天津市海河医院骨科
  • 收稿日期:2014-12-05 修回日期:2015-02-19 出版日期:2015-07-15 发布日期:2015-07-15
  • 通讯作者: 冯世庆 E-mail:fengsq@hotmail.com E-mail:fenfengou@hotmail.com
  • 作者简介:唐亮 （1982）， 男， 主治医师， 在职博士研究生在读， 主要从事脊柱脊髓损伤、 脊柱退行性疾病、 脊柱结核研究
  • 基金资助:
    天津市科学技术委员会科技计划项目 （08ZCKFSF03200）； 教育部新世纪人才支持计划 （NCET-06-0251）

Experimental study on repairing spinal cord injury by human umbilical cord blood CD34+ cell transplantation at different time points

TANG Liang, FENG Shiqing, GAO Ruixiao   

  1. 1 Tianjin Medical University General Hospital, Tianjin 300052, China; 2 Department of Orthopaedics, Tianjin Haihe Hospital
  • Received:2014-12-05 Revised:2015-02-19 Published:2015-07-15 Online:2015-07-15
  • Contact: FENG Shiqing E-mail:fengsq@hotmail.com E-mail:fenfengou@hotmail.com

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摘要： 目的 研究不同时期移植人脐血 CD34+细胞修复大鼠脊髓损伤的效果和机制。方法 免疫磁珠法从人新鲜脐血中分离得到 CD34+细胞。雌性 Wistar 大鼠 96 只， 以 IMPACTOR MODEL-Ⅱ脊髓损伤打击器建立 T10 脊髓损伤模型， 随机均分为免疫抑制剂应用组、 损伤后急性期移植组和损伤后亚急性期移植组， 对各组后肢功能恢复情况进行 BBB 评分， 损伤中心行双重免疫荧光染色、 2,3,5-氯化三苯基四氮唑(TTC)染色和血管明胶墨汁灌注观察。结果 损伤后第 8～56 天， 细胞急性期移植组的 BBB 评分高于其余 2 组(P < 0.05)； TTC 染色示组织活力降低区域比例小于其余 2 组(P < 0.01)； 明胶墨汁灌注示脊髓损伤中心血管密度大于其余 2 组(P < 0.01)； 亚急性期移植组的细胞存活密度大于急性期移植组（个/视野： 7.51±1.00 vs 5.51±0.89， t=6.051， P < 0.01）， 2 组均未观察到移植细胞的神经分化。结论 人脐血 CD34+细胞急性期移植可通过提高脊髓损伤中心血管密度促进微循环恢复， 增加组织活力，促进大鼠脊髓损伤后肢体功能恢复。

关键词: 脊髓损伤, 脐血干细胞移植, 人脐血 CD34+细胞, 组织活力, 血管密度, 神经分化

Abstract: Objective To investigate the effect of transplantation of human umbilical cord blood CD34+ cells on spinal cord injury. Methods CD34+ cells were separated from fresh human umbilical cord blood by magnetic cell sorting. Ninetysix female Wistar rats were injured at T10 by IMPACTOR MODEL-Ⅱ, and then randomly assigned to three groups: Cyclosporin A (CsA)+Dexamethasone (Dex) treated group (Ⅰ, n=32), local transplantation of cells+CsA+Dex treated group (Ⅱ) at the first day after operation (DAO 1, n=32), local transplantation of cells+ CsA+Dex treated group ( Ⅲ) at DAO 6 (n=32). BBB locomotor scoring system was used to assess the recovery of the lower limbs. The survival and neural differentiation of transplanted cells at the injury site were observed by double immunofluorescence. The tissue vitality at the injury site was observed by 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) staining, and the blood vessel density was observed by infusing mixture of Chinese ink and glutin followed by HE staining. Results BBB score at DAO 8-56 was significantly higher in Ⅱ group than that of other two groups (P < 0.05). TTC staining showed that the proportion of decreased vitality area was significantly smaller inⅡ group than that of other two groups (P < 0.01). The result of gelatin ink perfusion showed that the blood vessel density at the injury site was significantly bigger in Ⅱ group than that of other two groups (P < 0.01). There were more survival transplanted cells inⅡ group than those of III group (per visual field, 7.51±1.00 vs 5.51±0.89， t=6.051, P < 0.01). All the transplanted cells didn’ t differentiate into neural cells. Conclusion Human umbilical cord blood CD34+ cells can promote the recovery of the lower limbs after spinal cord injury by repairing blood vessels to increase tissue vitality at the injury site in rats.

Key words: spinal cord injury, cord blood stem cell transplantation, human umbilical cord blood CD34 + cells, tissue vitality, blood vessel density, neural differentiation