不同分化状态脐血间充质干细胞移植对脑梗死的影响(1)
【摘 要】目的 探讨不同分化状态脐血间充质干细胞(umbilical cord blood mesenchymal stem cells，UCB-MSCs)对脑梗死的治疗影响。方法 分离培养UCB-MSCs并构建大鼠脑梗死模型；利用脑立体定位仪经侧脑室分别注射原代培养的UCB-MSCs和经碱性成纤维细胞生长因子(basic fibroblast growth factor，bFGF)诱导培养的UCB-MSCs进行移植治疗大鼠脑梗死模型。免疫组化观察移植后的UCB-MSCs在脑梗死区分布及脑梗死周围组织胶质纤维酸性蛋白(glial fibrillary acidic protein，GFAP)与神经元特异性烯醇化酶(neuron-specific enolase，NSE)的表达；同时，在细胞移植后的3d、7d、14d和28d对大鼠进行神经功能评价。结果 诱导后UCB-MSCs组大鼠脑梗死灶周围的GFAP、NSE阳性细胞数量及其神经功能评分在7d、14d、28d时间点均高于未诱导组。结论 诱导后的UCB-MSCs移植能对脑梗死大鼠模型缺血脑组织起到修复作用，并能促进脑梗死后大鼠神经功能改善。
【关键词】脑梗死；脐血间充质干细胞；碱性成纤维细胞生长因子；诱导
Effects of differentiated umbilical cord blood mesenchymal stem cells on cerebral infarction
Abstract：objective To explore the effection of differentiated umbilical cord blood mesenchymal stem cells(UCB-MSCs)on the treatment of cerebral infarction.Methods To isolate and culture UCB-MSCs and construct cerebral infarction model rats；Using brain stereotaxic instrument，primary cultured UCB-MSCs and cultured UCB-MSCs induced by basic fibroblast growth factor(bFGF)were respectively injected and transplanted into the lateral ventricles of the cerebral infarction model rats.By immunohistochemistry the distribution of the transplanted UCB-MSCs and the expression of glial fibrillary acidic protein(GFAP)and neuron-specific enolase(NSE)were observed separately in the cerebral infarction area.At the same time，the nervous function of the cerebral infarction model rats was evaluated after 3 days、7 days、14 days and 28 days.Results The positive expression of GFAP and NSE around the cerebral infarction area and the neurological function score in 7d、14d and 28d time point of the induced group were higher than the non-induced group.Conclusion The induced UCB-MSCs can repair the ischemia brain tissues of the cerebral infarction model rats and improve neural function of rats after cerebral infarction.
Key words：cerebral infarction；umbilical cord blood mesenchymal stem cells；fibroblast growth factors；induce
引 言
脑梗死后大脑梗死区域神经细胞缺血缺氧，导致细胞不可逆的坏死，进一步导致受损区域支配的机体神经功能缺失，临床上表现为瘫痪、失语、痴呆、大小便失禁等严重后遗症，给家庭和社会带来了沉重的负担。目前，针对脑梗死患者临床上主要采取药物治疗、介入治疗和外科手术治疗等措施，但这些措施并不能从根本上挽救受损的神经细胞，进而最大程度提高治疗效果和改善预后。近年来，随着国内学术界对干细胞研究的逐渐重视与深入，研究学者提出干细胞移植将会成为促使脑梗死后神经功能恢复的新策略。本研究将经碱性成纤维细胞生长因子(bFGF)诱导后的UCB-MSCs和未经诱导的原代UCB-MSCs分别输入脑梗死的大鼠体内，通过大鼠神经功能评分和大脑免疫组织化学切片分析何种状态的UCB-MSCs疗效更佳，为临床干细胞移植治疗脑梗死提供理论依据。
方 法
1人UCB-MSCs的分离、培养与鉴定
1.1人UCB-MSCs的分离 无菌条件下取正常足月剖腹产胎儿的脐血40-80mL，20 U/mL肝素抗凝，采集后12 h内分离淋巴细胞；随后将新鲜脐带血与4℃1×PBS液等体积充分混匀、稀释，叠加于淋巴细胞分离液上(密度为1.077 g/mL)，室温以1500 r/ min离心15min；待离心管中液体分为4层，吸取血清层和离液层之间的白色云雾状有核细胞层，以4℃1×PBS稀释，800 r/min离心5 min，洗涤2次，少量培养基悬浮细胞、计数。, 百拇医药(韩小改1 牛朝霞2 单 泓2 李建斌1)
【关键词】脑梗死；脐血间充质干细胞；碱性成纤维细胞生长因子；诱导
Effects of differentiated umbilical cord blood mesenchymal stem cells on cerebral infarction
Abstract：objective To explore the effection of differentiated umbilical cord blood mesenchymal stem cells(UCB-MSCs)on the treatment of cerebral infarction.Methods To isolate and culture UCB-MSCs and construct cerebral infarction model rats；Using brain stereotaxic instrument，primary cultured UCB-MSCs and cultured UCB-MSCs induced by basic fibroblast growth factor(bFGF)were respectively injected and transplanted into the lateral ventricles of the cerebral infarction model rats.By immunohistochemistry the distribution of the transplanted UCB-MSCs and the expression of glial fibrillary acidic protein(GFAP)and neuron-specific enolase(NSE)were observed separately in the cerebral infarction area.At the same time，the nervous function of the cerebral infarction model rats was evaluated after 3 days、7 days、14 days and 28 days.Results The positive expression of GFAP and NSE around the cerebral infarction area and the neurological function score in 7d、14d and 28d time point of the induced group were higher than the non-induced group.Conclusion The induced UCB-MSCs can repair the ischemia brain tissues of the cerebral infarction model rats and improve neural function of rats after cerebral infarction.
Key words：cerebral infarction；umbilical cord blood mesenchymal stem cells；fibroblast growth factors；induce
引 言
脑梗死后大脑梗死区域神经细胞缺血缺氧，导致细胞不可逆的坏死，进一步导致受损区域支配的机体神经功能缺失，临床上表现为瘫痪、失语、痴呆、大小便失禁等严重后遗症，给家庭和社会带来了沉重的负担。目前，针对脑梗死患者临床上主要采取药物治疗、介入治疗和外科手术治疗等措施，但这些措施并不能从根本上挽救受损的神经细胞，进而最大程度提高治疗效果和改善预后。近年来，随着国内学术界对干细胞研究的逐渐重视与深入，研究学者提出干细胞移植将会成为促使脑梗死后神经功能恢复的新策略。本研究将经碱性成纤维细胞生长因子(bFGF)诱导后的UCB-MSCs和未经诱导的原代UCB-MSCs分别输入脑梗死的大鼠体内，通过大鼠神经功能评分和大脑免疫组织化学切片分析何种状态的UCB-MSCs疗效更佳，为临床干细胞移植治疗脑梗死提供理论依据。
方 法
1人UCB-MSCs的分离、培养与鉴定
1.1人UCB-MSCs的分离 无菌条件下取正常足月剖腹产胎儿的脐血40-80mL，20 U/mL肝素抗凝，采集后12 h内分离淋巴细胞；随后将新鲜脐带血与4℃1×PBS液等体积充分混匀、稀释，叠加于淋巴细胞分离液上(密度为1.077 g/mL)，室温以1500 r/ min离心15min；待离心管中液体分为4层，吸取血清层和离液层之间的白色云雾状有核细胞层，以4℃1×PBS稀释，800 r/min离心5 min，洗涤2次，少量培养基悬浮细胞、计数。, 百拇医药(韩小改1 牛朝霞2 单 泓2 李建斌1)