小鼠胎肝间充质干细胞移植治疗缺血性脑损伤的实验研究  

Transplantation treatment of ischemic CNS injury in mice with mesenchymal stem cells from murine fetal liver

作　　者：孙艳[1,2] 张洹[1] 何冬梅[1] 

机构地区：[1]暨南大学医学院血液病研究所,510632 [2]广东药学院生命科学与生物制药学院

出　　处：《中国康复医学杂志》2009年第2期142-145,I0002,共5页Chinese Journal of Rehabilitation Medicine

基　　金：国务院侨办重点学科建设基金(2005)

摘　　要：目的:探讨小鼠胎肝间充质干细胞(flMSCs)移植对缺血性脑损伤是否具有修复治疗作用。方法:用快速PCR扩增Y染色体特异sry基因方法鉴定并分离BALB/c雄性小鼠flMSCs,将其植入雌性缺血性脑损伤BALB/c小鼠损伤侧侧脑室。40d后观察神经功能,PCR检测脑组织sry基因;HE染色和尼氏染色观察脑组织结构;免疫组化和FISH双染观察flMSCs分化存活。结果:5只移植小鼠长期存活;移植后40 d,小鼠神经功能缺损有所改善;脑组织结构未见明显异常;移植小鼠脑组织sry基因阳性;(6.6±2.9)%的受体鼠损伤侧脑细胞Y染色体阳性,Y染色体和Nestin双阳性细胞占缺血侧脑组织细胞的(1.1±0.9)%,Y染色体和Tubulin双阳性细胞占(3.2±1.4)%,Y染色体和GFAP双阳性细胞占(0.9±0.7)%,对侧脑组织未见到染色体阳性细胞。结论:缺血性脑损伤条件下,小鼠flMSCs可以向神经细胞分化,参与损伤脑组织的结构重建。Objective： To investigate whether murine fetal liver mesenchymal stem cells （flMSCs） could help to repair CNS injury and recover the neural function in ischemic isogenic mouse. Method： Sex of BALB/c mouse fetuses was determined by rapid PCR analysis of sty gene on Y chromosome. Male flMSCs were isolated and injected into the lateral ventricle of injured hemisphere in female mice with ischemia. Recovery of neurological deficit was reevaluated at 40 days after transplantation. HE stain and Nissl stain were employed to observe pathological change in injured brain. Whether the flMSCs grafts were survival was determined by PCR assay of mouse sry gene in recipient＇s brain. The donor cells and their differentiation to neural cells were detected and identified by FISH and immunohistochemistry double-staining analysis. Result： Five of those mice received flMSCs grafts survived and had no problem with normal action. Neurological deficit rehabilitations were observed after 40 days. Brain tissues stained by HE and Nissel didn＇t show significant abnormality. PCR assay detected sry gene in brains of recipients. The ratio of Y chromosome-positive cells in injured hemisphere was （6.6±2.9）%. Ratio of both Y chromosome-positive cells and Nestin positive cells was （1.1±0.9）%. Ratio of both Y chromosome-positive ceils and Tubulin positive cells was（3.2±.4）%. Ratio of both Y chromosome-positive cells and GFAP positive cells was （0.9±0.7）%. No Y chromosome-positive cell was found in contralateral hemisphere. Conclusion： Murine flMSCs could differentiate into neurocytes in ischemic injured CNS and might be helpful for reconstitution of injured hemisphere.

关 键 词：小鼠 胎肝 间充质干细胞 脑缺血 Y染色体 

分 类 号：R743.3[医药卫生—神经病学与精神病学]