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国家自然科学基金(30500517)

作品数:2 被引量:33H指数:2
相关作者:高爱丽梁鹏刘恩重梁洪生徐小平更多>>
相关机构:东北农业大学哈尔滨医科大学更多>>
发文基金:国家自然科学基金哈尔滨市科技创新人才研究专项资金黑龙江省青年科学基金更多>>
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Human neural stem cells promote corticospinal axons regeneration and synapse reformation in injured spinal cord of rats被引量:31
2006年
Background Axonal regeneration in lesioned mammalian central nervous system is abortive, and this causes permanent disabilities in individuals with spinal cord injuries. This paper studied the action of neural stem cell (NSC) in promoting corticospinal axons regeneration and synapse reformation in rats with injured spinal cord. Methods NSCs were isolated from the cortical tissue of spontaneous aborted human fetuses in accordance with the ethical request. The cells were discarded from the NSC culture to acquire NSC-conditioned medium. Sixty adult Wistar rats were randomly divided into four groups (n=15 in each): NSC graft, NSC medium, graft control and medium control groups. Microsurgical transection of the spinal cord was performed in all the rats at the T11. The NSC graft group received stereotaxic injections of NSCs suspension into both the spinal cord stumps immediately after transection; graft control group received DMEM injection. In NSC medium group, NSC-conditioned medium was administered into the spinal cord every week; NSC culture medium was administered to the medium control group. Hindlimb motor function was assessed using the BBB Locomotor Rating Scale. Regeneration of biotin dextran amine (BDA) labeled corticospinal tract was assessed. Differentiation of NSCs and the expression of synaptophysin at the distal end of the injured spinal cord were observed under a confocal microscope. Group comparisons of behavioral data were analyzed with ANOVA. Results NSCs transplantation resulted in extensive growth of corticospinal axons and locomotor recovery in adult rats after complete spinal cord transection, the mean BBB scores reached 12.5 in NSC graft group and 2.5 in graft control group (P〈 0.05). There was also significant difference in BBB score between the NSC medium (11.7) and medium control groups (3.7, P〈 0.05). BDA traces regenerated fibers sprouted across the lesion site and entered the caudal part of the spinal cord. Synaptophysin expression colocalized with BDA positiv
LIANG PengJIN Lian-hongLIANG TaoLIU En-zhongShi-guang
新型联合基质桥接大鼠脊髓横断性损伤断端促进运动功能恢复实验研究被引量:2
2009年
目的研究新型联合基质——BMSCs-去上皮细胞羊膜(denuded human amniotic membrane,DHAM)桥接大鼠脊髓横断性损伤断端,促进轴突再生及提高后肢运动功能的作用。方法取健康产妇正常剖宫产术后自愿捐献的羊膜,经联合消化法彻底去除表面上皮细胞制备DHAM,并行HE染色观察。健康4周龄雌性Wistar大鼠4只,取骨髓行BMSCs分离、培养,取第4代细胞以PKH26标记后,以1×105个/cm2密度接种于DHAM上制备联合基质(BMSCs-DHAM)。倒置相差显微镜和激光共聚焦显微镜观察BMSCs生长状态。另取8周龄雌性Wistar大鼠40只,体重200~220g,制备T9脊髓横断性损伤模型,随机分为4组,每组10只。A、C组分别用BMSCs-DHAM和单纯DHAM包裹脊髓的2个断端,B、D组分别于脊髓损伤中心注射10μL细胞浓度为1×104个/μL的BMSCs和等量生理盐水。术后12周采用BBB运动评分检测大鼠后肢运动功能恢复情况,行皮层运动诱发电位(motion evoked potential,MEP)测定;生物素化葡聚糖胺(biopinylated dext anamine,BDA)顺行示踪检测及神经丝蛋白200(neurofilament protein200,NF-200)免疫荧光染色观察。结果HE染色显示联合消化法成功制备DHAM。镜下观察到BMSCs在DHAM上生长状态良好。术后12周,A组BBB评分达(12.50±1.26)分,明显高于其余各组(P<0.05)。MEP测定显示,A组潜伏期(3.52±2.45)ms及波幅(480.68±18.41)μV,恢复效果优于其余各组(P<0.05)。BDA顺行示踪观察示,A组损伤平面以下BDA通过损伤部位的轴突再生率为54.12%±3.30%,均高于其余各组(P<0.05);且A组NF-200免疫荧光染色亦可见大量再生轴突通过损伤部位。结论新型联合基质BMSCs-DHAM一定程度上提高了脊髓横断性损伤大鼠的后肢运动功能,是一种具有广泛应用前景的基质材料。
梁洪生梁鹏高爱丽祁权刘恩重吴佳宁徐小平
关键词:BMSCS轴突再生
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