摘要
目的探讨脑内移植人脐血间充质干细胞(MSCs)对新生大鼠缺氧缺血性脑损伤(HIBD)的治疗效果。方法无菌条件下收集20份健康足月新生儿脐血,采用密度梯度离心法分离出脐血单个核细胞,再通过贴壁细胞培养法培养出人脐血MSCs。选择P3代人脐血MSCs作为移植细胞,并经5-溴尿嘧啶脱氧核苷(BrdU)体外标记。7日龄SD大鼠30只制备HIBD模型,造模过程中死亡1只,剩余29只分为移植组(n=18)和对照组(n=11)。造模后第3天,在立体定位条件下,移植组大鼠经左侧大脑皮层注入人脐血MSCs,对照组在相同部位注入相同体积的PBS。在细胞移植后第7天,随机选择6只移植组大鼠处死取脑,通过脑组织免疫组织化学检测方法观察移植细胞在脑内的存活、迁移和分化情况。于移植后第1、7、14、21、28天采用改良神经功能损害评分(mNSS)方法对二组大鼠的神经功能进行评价。结果20份足月新生儿脐血中5份培养出人脐血MSCs,培养成功率为25%。移植组大鼠脑组织免疫组织化学检测发现移植细胞在脑内能够存活,并以移植点为中心向周围迁移,其中(12.67±2.73)%的移植细胞分化为星形胶质细胞样细胞,未发现移植细胞向神经元样细胞分化。mNSS检测结果显示,移植第1、7天移植组大鼠mNSS低于对照组,但差异无显著性意义(Pa>0.05),第14、21、28天移植组大鼠mNSS低于对照组,差异有显著性意义(Pa<0.05)。结论人脐血MSCs脑内移植对新生大鼠HIBD具有较好的治疗作用。
Objective To investigate the therapeutic effect of intracerebral transplantation of mesenchymal stem ceils（MSCs） derived from human umbilical cord blood（UCB） on hypoxic - ischemic brain damage（HIBD） in neonatal rat. Methods Twenty samples of human UCB were collected from healthy full - term newborns. MSCs were isolated from human UCB by density gradient centrifugation and purified by adhere cell selection method. For transplantation, P3 human UCB - derived MSCs were labeled by the 5 - bromo - 2 - deoxyuridine （BrdU）. Thirty SD rats of 7 d were built for neonatal HIBD model. One rat died and others were divided into transplant group（ n = 18 ） and control group（ n = 11 ）. At the third day after building models,human UCB -derived MSCs were injected into left cortex in transplant group, while PBS of the same volume was injected into the same site in control group at the same time. The seventh day after transplantation,6 rats of trans- plant group were sacrificed to prepare brain tissue sections. The survival, migration and differentiation of the transplanted cells were investiga- ted by brain tissue immunohistochemical analysis, and nervous function of 2 groups were evaluated by modified neurological severity score （mNSS） on the first,7th ,14th ,21th and 28th day after transplantation. Results MSCs were isolated from 5 of 20 human UCB samples. Immunocytochemical analysis of brain tissue showed that the transplanted human UCB - derived MSCs could survive and migrate around by the cen- ter of transplant site. There were （ 12.67 ± 2.73 ） % of MSCs differentiated into astrocyte - like cells, mNSS showed that the score of transplant group was lower than that of control group on the first,7th, 14th ,21th and 28th day,and the differences of score points between 2 groups on the 14th ,21 th and 28th day were statistically significant（ Pa 〈 0.05 ）. Conclusion It is effective for the transplantation of human UCB - derived MSCs to treat neonatal rat HIBD. J Appl Clin Pediatr, 2008,23 （14） ： 1090 - 1092
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2008年第14期1090-1092,共3页
Journal of Applied Clinical Pediatrics
基金
福建省自然科学基金项目资助(C0310036)
关键词
间充质干细胞
人脐血
脑损伤
缺氧缺血性
移植
mesenchymal stem cells
human umbilical cord blood
hypoxic - ischemic brain damage
transplant
