脊髓损伤亚急性期移植神经干细胞：损伤脊髓5-羟色胺的表达☆

背景：5-羟色胺能纤维可以作为评价脊髓损伤后再生的指标之一。 目的：观察神经干细胞在脊髓损伤的亚急性期(第7天)脊髓内移植对大鼠受损伤脊髓再生修复的影响。 方法：将正常成年SD雌性大鼠分为3组,单纯脊髓全横断组、假手术组(仅仅打开椎板,但不横断脊髓)、神经干细胞移植组。神经干细胞移植组在脊髓全横断后第7天时进行神经干细胞移植,其他两组不移植神经干细胞。 结果与结论：神经干细胞移植组瘢痕上1至2节段5-羟色胺的阳性纤维数量多于单纯脊髓全横断组。提示神经干细胞亚急性期移植能部分促进脊髓损伤后脊髓神经的再生。 关键词：脊髓损伤;再生;神经干细胞;5-羟色胺;大鼠 doi:10.3969/j.issn.1673-8225.2012.06.023     

神经干细胞移植治疗脊髓损伤

背景：研究已证实神经干细胞能促进脊髓损伤大鼠神经功能的恢复,但对移植细胞在体内的增殖、分化、迁移的研究有限。 目的：观察神经干细胞移植对脊髓损伤大鼠后肢运动功能修复的影响。 方法：SD大鼠制成T10脊髓全横断损伤模型,于造模成功后1周采用局部微量注射法。随机数字表法分为3组：损伤对照组仅打开椎管暴露脊髓;移植对照组：注射10 μL DMEM/F12培养液;细胞移植组：造模后移植浓度为1.0×109 L-1的神经干细胞悬液10 μL。移植后通过不同时间点BBB行为评分、病理组织学、免疫荧光技术评价大鼠脊髓功能修复情况及移植细胞在体内的存活、迁移、分化。 结果与结论：在体外成功建立SD大鼠海马源性神经干细胞培养体系;移植对照组、细胞移植组大鼠随着时间延长BBB评分均不同程度提高,从移植后2周起细胞移植组大鼠评分明显高于移植对照组(P < 0.05);神经干细胞移植后能够在体内继续存活、迁移并且分化为NF-200、GFAP表达阳性的神经元及星形胶质细胞。提示神经干细胞移植治疗脊髓损伤是一种有效的方法。     

补阳还五汤对大鼠脊髓损伤后移植神经干细胞存活、增殖与迁移的影响

目的:探讨补阳还五汤(Buyang Huanwu Decoction,BYHWD)灌胃对脊髓损伤(spinal cord injury,SCI)后移植胚胎神经干细胞(neural stem cells,NSCs)存活、增殖与迁移的影响。方法:将SD大鼠随机分为SCI组、NSCs移植组、NSCs移植联合补阳还五汤灌胃组。制作大鼠脊髓横断损伤模型,将BrdU标记NSCs细胞按浓度1×106/ml分别移植入移植组和联合组的脊髓横断处,损伤组用等量DMEM/F12完全培养基代替。术后联合组每天用BYHWD予以灌胃治疗1次,损伤组和移植组用生理盐水代替。各组在7、14、28 d后分别取材,用免疫荧光组织化学法检测移植在SCI处的NSCs存活、增殖和迁移情况。结果:损伤组未见BrdU标记阳性细胞;联合组各时间点BrdU标记阳性细胞数量均多于移植组各时间点,并有显著性统计学意义(P<0.05);联合组28 d时BrdU标记阳性细胞数量最多,与联合组7 d和14 d相比有显著性统计学意义(P<0.05);在联合组各时间点,BrdU标记阳性细胞向SCI处头尾端组织迁移的距离明显长于移植组各时间点。结论:脊髓横断损伤后,移植胚胎NSCs能够存活并增殖和迁移。补阳还五汤能够促进大鼠脊髓全横断损伤处移植的NSCs存活、增殖和迁移。     

神经干细胞移植对小鼠脊髓损伤的影响

目的观察神经干细胞(NSCs)移植对脊髓损伤(SCI)小鼠功能恢复的影响。方法分离、培养、增殖和纯化E14-17d小鼠的NSCs,并通过免疫荧光法对其进行鉴定。将绿色荧光蛋白转基因小鼠的NSCs移植到小鼠脊髓损伤模型体内,术后进行行为学和病理学检测,观察小鼠功能恢复情况。运用改良Allen's法制备小鼠T10-T11脊髓损伤动物模型,动物分为假手术组(12只)、模型组(12只),治疗组(12只)和对照组6(12只)。治疗组每只小鼠自眶静脉注射NSCs悬液200μl(2×10个细胞),对照组只注射DMEM/F12培养基200μl。术后1d、3d、7d、14d、21d、28d和56d,进行BBB运动功能评分和病理学检测,观察植入区细胞生长变化情况。结果 BBB评分显示:治疗组明显高于对照组(<0.01),治疗组与假手术组相比没有明显差异(>0.05),说明NSCs移植后小鼠的行为学得到了明显改善,功能有所恢复。病理学检测发现,移植后NSCs不仅迁移到脊髓损伤区,而且与宿主细胞较好地整合。结论移植的E14-17d胚胎小鼠NSCs不仅可在脊髓损伤部位存活并和宿主细胞整合,而且可促进小鼠后肢运动功能恢复。     

Transplantation of Schwann cells to subarachnoid space induces repair in contused rat spinal cord

Schwann cell transplantation is well known to induce repair in the injured spinal cord which disables millions of injured patients throughout the world. An ideal route of delivering the grafted Schwann cells to the spinal cord should neither cause more injury nor reinitiate inflammatory events and also provide a favorable milieu to the grafted cells. In this study, we have utilized subarachnoid route to transplant Schwann cells and evaluated their effects in a contusive model of spinal cord injury. Adult rats weighing 100-140 g were experimentally injured by crushing the spinal cord with a titanium clip and then divided into four groups (Tracing, Control, Medium-treated and Schwann cell-treated). Cultured Schwann cells (5x10(4) cells in 5 microl) or medium were injected to the animals of corresponding groups via subarachnoid space at the injured site 7 days after injury. In tracing group, Schwann cells (labeled with Hoechst) demonstrated their presence within spinal cord 7 days after transplantation. Evaluation of locomotor performance of animals for 60 days after injury showed that animals treated with Schwann cells had significant improvement (P<0.01). Similarly, the axon density at the site of injury was significantly higher. The results indicate the efficacy of subarachnoid route for the transplantation of Schwann cells in inducing repair of the contused spinal cord. We conclude that this route can be useful for the transplantation of Schwann cells and offers a hope for the patients suffering from spinal cord injury.     

骨髓间充质干细胞向脊髓损伤区的迁徙

背景：骨髓间充质干细胞移植到脊髓损伤区域后,如何观察其在体内的生存和转归情况,一直是让人困扰的问题。 目的：观察骨髓间充质干细胞在大鼠脊髓损伤区内的迁徙情况。 方法：36只Wistar大鼠随机分为2组,实验组制作脊髓损伤模型1周后,经尾静脉移植用DAPI标记的骨髓间充质干细胞(1×109 L-1) 1 mL,连续注射2 d。对照组未行脊髓损伤,与实验组同一时间同法行骨髓间充质干细胞移植。分别于移植后5,10,15 d,制作损伤脊髓冰冻切片,在激光共聚焦显微镜下观察骨髓间充质干细胞的迁徙情况。 结果与结论：实验组于移植后5 d,在脊髓损伤组织血管内出现少量荧光标记的骨髓间充质干细胞,10 d后有血管外弥散,15 d后有广泛弥散。对照组均未见DAPI标记的骨髓间充质干细胞。结果表明骨髓间充质干细胞经大鼠尾静脉移植后,能透过血脊髓屏障向损伤脊髓组织迁徙。     

促红细胞生成素促进脊髓损伤大鼠移植神经干细胞存活和迁移

背景：如何有效促进移植入脊髓损伤组织内的神经干细胞存活和迁移,是目前神经修复研究的重点。 目的：观察促红细胞生成素对脊髓损伤大鼠移植神经干细胞存活、增殖和迁移的影响。 方法：将60只SD大鼠随机分为3组,均制备脊髓横断损伤模型。造模7 d,神经干细胞移植组和促红细胞生成素组于脊髓损伤处移植BrdU标记的神经干细胞7 μL(1×10⁹ L^-1),脊髓损伤对照组移植DMEM/F12培养基;促红细胞生成素组腹腔内注射促红细胞生成素5 000 U/kg,1次/d,连续注射7 d,其余两组注射等量生理盐水。于细胞移植后8周取损伤脊髓组织。 结果与结论：造模2周后,神经干细胞移植组和促红细胞生成素组BBB评分明显高于脊髓损伤对照组(P < 0.05),造模4周后,促红细胞生成素组BBB评分明显高于神经干细胞移植组(P < 0.05)。免疫荧光染色显示促红细胞生成素组大鼠损伤脊髓组织BrdU阳性细胞数量及迁移距离均大于神经干细胞移植组(P < 0.05)。说明促红细胞生成素能促进损伤脊髓组织原位移植的神经干细胞的存活与迁移,加速神经功能修复。     

Intravenously injected neural progenitor cells of transgenic rats can migrate to the injured spinal cord and differentiate into neurons, astrocytes and oligodendrocytes

Transplantation of neural progenitor cells (NPCs) has been reported recently to promote regeneration of the injured spinal cord. In the majority of these reports, cell transplantation was performed by local injection with a needle. However, direct injection might be too invasive for clinical use; therefore, the authors investigated a new method of delivering NPCs for the treatment of spinal cord injury. In this study, NPCs were obtained from E15 fetal hippocampus of transgenic rats expressing green fluorescent protein and 100,000 cells were transplanted intravenously into each animal 24h after contusion injury. It was found that the injected NPCs migrated to the lesion site widely and demonstrated nestin at an early phase after transplantation. These NPCs differentiated into neurons, astrocytes and oligodendrocytes, and survived at least for 56 days. These results indicated that intravenously injected neural stem cells migrated into the spinal cord lesion while preserving their potential as NPCs, and that this procedure is a potential method of delivering cells into the lesion for the treatment of spinal cord injury.     

不同时间点静脉移植嗅鞘细胞修复脊髓损伤

背景:嗅鞘细胞具有促进轴突再生、为受伤的宿主细胞提供营养支持以及调节炎症反应的能力,是修复脊髓损伤具有潜力的细胞。目的:探讨通过静脉移植嗅鞘细胞治疗脊髓损伤的最佳移植时间窗。方法:SPF雄性SD大鼠30只,采用脊髓半横断建立大鼠脊髓损伤模型,并随机分为5组:脊髓损伤后注射嗅鞘细胞1 d移植组、3 d移植组、7 d移植组、10 d移植组及PBS对照组。应用荧光量子点标记嗅鞘细胞;分别于1,3,7,10 d时间点通过尾静脉移植应用量子点标记的嗅鞘细胞;PBS对照组脊髓损伤后注射PBS。注射后1 d取损伤处的脊髓;应用小动物成像仪测定不同时间点转移到损伤处荧光的数值,通过荧光的强度衡量转移到损伤处细胞的量;应用嗅鞘细胞Anti-p75 NGF Receptor抗体做损伤处脊髓的免疫组织化学。实验方案经宁夏医科大学动物实验伦理委员会批准(编号:2017-073)。结果与结论:①荧光量子点可以标记嗅鞘细胞;②荧光测定结果与免疫组织化学染色结果:1,3,7,10 d时间点通过尾静脉移植的嗅鞘细胞均有细胞转移到损伤处,7 d移植的嗅鞘细胞转移到损伤处的最多;③结果说明,脊髓损伤不同时间点注射的嗅鞘细胞均可以转移到脊髓损伤处,损伤后7 d移植的嗅鞘细胞转移到损伤处的细胞数量最多,可以作为移植的最佳时间窗。     

Long-term Fate of Allogeneic Neural Stem Cells Following Transplantation into Injured Spinal Cord

To characterize the fate of allogeneic neural stem cells (NSCs) following transplantation into injured spinal cord, green fluorescent protein (GFP)-NSCs isolated from GFP transgenic Sprague–Dawley rat embryos were transplanted into contused spinal cords of Wistar rats. The GFP-NSCs survived for at least 6 months in injured spinal cord; most of them differentiated rapidly into astrocytes, and a few were able to undergo proliferation. After transplantation, the GFP-NSCs remained in the transplantation site at the early stage, and then migrated along white-matter, and gathered around the injured cavity. At 6 months post-transplantation, CD8 T-lymphocytes infiltrated the spinal cord, and mixed lymphocyte culture from host and donor showed that lymphocytes from the host spleen were primed by allogeneic GFP-NSCs. At 12 months post-transplantation, most GFP cells in the spinal cord lost their morphology and disintegrated. The Basso, Beattie and Bresnahan score and footprint analysis indicated that the improvement of locomotor function in transplanted rats appeared only at the early stage, and was not seen even at 6 months after transplantation All these results suggest that the allogeneic NSCs, after transplantation into injured spinal cord, activate the host immune system. Therefore, if immunosuppressive agents are not used, the grafted allogeneic NSCs, although they can survive for a long time, are subjected to host immune rejection, and the effect of NSCs on functional recovery is limited.     

直流电场诱导神经干细胞桥接横断脊髓的实验研究

目的:探讨直流电场(direct current field,DC)诱导神经干细胞(neural stem cells,NSCs)桥接横断脊髓的作用。方法:将54只雌性成年SD大鼠随机分为脊髓损伤(spinal cord injury,SCI)组、NSCs移植组、NSCs移植联合直流电场诱导组(DC+NSCs组),各组分7,14,28d三个时间点取材,HE染色观察神经纤维再生情况,透射电镜下观察脊髓白质内髓鞘结构变化,免疫荧光染色检测髓磷脂碱性蛋白(myelin basic protein,MBP)表达变化。结果:在光镜下,DC+NSCs组神经纤维较其他两组都多,排布均匀且走行较规则,部分纤维穿越损伤脊髓断端两侧。在电镜下,DC+NSCs组髓鞘松散程度明显减轻,部分髓鞘结构规整,排列有序,线粒体肿胀减轻,微管空泡样变较轻,可发现大量突触,薄髓的再生髓鞘明显增多。DC+NSCs组大鼠脊髓组织中可见直径大小不等的MBP阳性的圆形结构,MBP阳性表达最多,较其他两组有显著性差异(P0.01)。结论:直流电场诱导NSCs治疗SCI可以促进损伤处脱髓的轴突再髓鞘化和MBP的表达,促进神经纤维再生,从而起到桥接横断脊髓的作用。     

移植胚鼠神经干细胞对大鼠脊髓损伤后红核神经元损伤的影响

目的研究神经干细胞(NSCs)移植对大鼠脊髓损伤(SCI)后红核神经元的作用。方法5-溴脱氧尿嘧啶核苷(BrdU)法标记处于对数生长期的NSCs,采用电控脊髓损伤打击装置制作大鼠脊髓损伤模型。实验分为3组:NSCs组、SCI组和假手术组(Sham组)。SCI后3 d进行NSCs移植,用免疫组化法观察移植细胞的存活及迁移情况,用辣根过氧化物酶(HRP)逆行示踪技术标记红核神经元,并用四甲基联苯胺(TMB)呈色反应显示红核脊髓束神经元的存活情况,用行为学(BBB)评分法观察大鼠瘫痪肢体的恢复情况。结果在损伤脊髓区域可检测到BrdU标记的阳性NSCs,中脑HRP标记红核神经元数目明显多于SCI组(P<0.01),BBB评分亦明显高于SCI组(P<0.01)。结论体外培养的胚胎大鼠NSCs在移植到脊髓损伤区域后可存活和迁移,对SCI后中脑红核神经元具有保护作用,从而促进了大鼠肢体功能的恢复。     

神经干细胞移植修复大鼠脊髓半切伤的研究

目的：观察神经干细胞移植修复脊髓半切伤的疗效并探讨移植最佳时机。方法：制备大鼠T13-L1脊髓半切伤模型，取胎龄13、5d胎鼠大脑组织，体外分离、培养、诱导神经干细胞，并采用免疫细胞化学技术分别检测NSC（neural stemcell）特征性标志Nestin（巢蛋白）表达和用血清诱导分化为大量神经元NSE（neuron specific enolas）和神经胶质细胞GFAP（glial fibrillary acidic protein）表达。用明胶海绵吸附BrdU（5-溴脱氧尿嘧啶）标记好的神经干细胞悬液移植到脊髓半横断处，移植时间选择损伤后立即移植、损伤后第9、14d。观察移植后的大鼠行为的变化，通过CBS（combine behavioral score）评分和爬网格实验评价大鼠的运动功能恢复情况，并进行免疫组化鉴定，光学显微镜观察移植神经干细胞的存活和迁移。结果：在上述条件下培养及传代的细胞不断分裂增殖，形成悬浮生长的呈Nestin阳性的神经球；用血清诱导分化为大量表达NSE阳性的神经元和GFAP阳性的神经胶质细胞。与对照组相比，神经干细胞移植组明显修复了损伤结构，改善了下肢的功能，尤其是第9d移植组。结论：神经干细胞移植促进了脊髓损伤后神经结构和功能的恢复，是治疗脊髓半切伤一种有效方案，考虑综合因素移植干细胞的最佳时机应选损伤后9d左右。     

督脉电针对移植在大鼠脊髓全横断损伤处的神经干细胞存活分化和迁移的影响

目的探讨督脉电针对脊髓全横断损伤处移植的神经干细胞存活、分化和迁移的影响。方法将20只成年SD大鼠分为神经干细胞移植14d组（NSCs14d组）、督脉电针＋神经干细胞移植14d组（电针NSCs14d组）、神经干细胞移植30d组（NSCs30d组）和督脉电针＋神经干细胞移植30d组（电针NSCs30d组）4组。所有动物均实施T10段脊髓全横断手术,其中电针组和电针NSCs组于术后5d进行电针治疗。分别于术后14d和30d取材检测移植在脊髓损伤处的神经干细胞存活、分化和迁移情况。结果1．电针NSCs14d组或电针NSCs30d组移植的神经干细胞存活数量均多于NSCs14d组或NSCs30d组,但是电针NSCs30d组或NSCs30d组移植的神经干细胞存活数量均少于电针NSCs14d组或NSCs14d组。2．电针NSCs30d组和NSCs30d组的脊髓全横断损伤处及其相邻的组织均有少量移植的神经干细胞呈现微管相关蛋白2（MAP2）阳性染色。3．电针NSCs30d组和NSCs30d组的脊髓全横断损伤处及其相邻的组织均可观察到较多移植的神经干细胞呈现胶质原纤维酸性蛋白（GFAP）阳性染色。4．在电针NSCs14d组或电针NSCs30d组,移植的神经干细胞向脊髓损伤处尾端组织迁移的距离明显长于NSCs14d组或NSCs30d组。结论督脉电针能够促进大鼠脊髓全横断损伤处移植的神经干细胞存活,这些细胞能分化为MAP2或GFAP阳性细胞;督脉电针对移植在脊髓损伤处的神经干细胞向宿主脊髓组织迁移方向有一定的影响。     

Transplantation of human embryonic stem cell-derived oligodendrocyte progenitors into rat spinal cord injuries does not cause harm

Demyelination contributes to loss of function following spinal cord injury. We have shown previously that transplantation of human embryonic stem cell-derived oligodendrocyte progenitors into adult rat 200 kD contusive spinal cord injury sites enhances remyelination and promotes recovery of motor function. Previous studies using oligodendrocyte lineage cells have noted a correlation between the presence of demyelinating pathology and the survival and migration rate of the transplanted cells. The present study compared the survival and migration of human embryonic stem cell-derived oligodendrocyte progenitors injected 7 days after a 200 or 50 kD contusive spinal cord injury, as well as the locomotor outcome of transplantation. Our findings indicate that a 200 kD spinal cord injury induces extensive demyelination, whereas a 50 kD spinal cord injury induces no detectable demyelination. Cells transplanted into the 200 kD injury group survived, migrated, and resulted in robust remyelination, replicating our previous studies. In contrast, cells transplanted into the 50 kD injury group survived, exhibited limited migration, and failed to induce remyelination as demyelination in this injury group was absent. Animals that received a 50 kD injury displayed only a transient decline in locomotor function as a result of the injury. Importantly, human embryonic stem cell-derived oligodendrocyte progenitor transplants into the 50 kD injury group did not cause a further decline in locomotion. Our studies highlight the importance of a demyelinating pathology as a prerequisite for the function of transplanted myelinogenic cells. In addition, our results indicate that transplantation of human embryonic stem cell-derived oligodendrocyte progenitor cells into the injured spinal cord is not associated with a decline in locomotor function.     

Combined transplantation of neural stem cells and olfactory ensheathing cells for the repair of spinal cord injuries

Spinal cord repair is a problem that has long puzzled neuroscientists. The failure of the spinal cord to regenerate and undergo reconstruction after spinal cord injury (SCI) can be attributed to secondary axonal demyelination and neuronal death followed by cyst formation and infarction as well as to the nature of the injury environment, which promotes glial scar formation. Cellular replacement and axon guidance are both necessary for SCI repair. Multipotent neural stem cells (NSCs) have the potential to differentiate into both neuronal and glial cells and are, therefore, likely candidates for cell replacement therapy following SCI. However, NSC transplantation alone is not sufficient for spinal cord repair because the majority of the NSCs engrafted into the spinal cord have been shown to differentiate with a phenotype which is restricted to glial lineages, further promoting glial scaring. Olfactory ensheathing cells (OECs) are a unique type of glial cell that occur both peripherally and centrally along the olfactory nerve. The ability of olfactory neurons to grow axons in the mature central nervous system (CNS) milieu has been attributed to the presence of OECs. It has been shown that transplanted OECs are capable of migrating into and through astrocytic scars and thereby facilitating axonal regrowth through an injury barrier. Given the complementary properties of NSCs and OECs, we predict that the co-transplantation of NSCs and OECs into an injured spinal cord would have a synergistic effect, promoting neural regeneration and functional reconstruction. The lost neurocytes would be replaced by NSCs, while the OECs would build "bridges" crossing the glial scaring that conduct axon elongation and promote myelinization simultaneously. Furthermore, the two types of cells could first be seeded into a bioactive scaffold and then the cell seeded construct could be implanted into the defect site. We believe that this type of treatment would lead to improved neural regeneration and functional reconstruction after SCI.     

Single neural progenitor cells derived from EGFP expressing mice is useful after spinal cord injury in mice

Neural stem cells (NSCs) were widely used for studying the cell's replacement after transplantation in nervous system because of its specific characteristics. However, Stracing the cells after transplantation was still a problem. In the present study, we isolated and cultured the neural stem cells from the C57BL/6J EGFP transgenic mouse (EGFP mice), and identified the capacity for self-renewal and differentiation into the three CNS lineages (neurons, astrocytes, and oligodendrocytes). Then we transplanted the single neural stem cell into the lesion spinal cord. Expression of GFP and differentiation was evaluated at two weeks post-transplantation. The data showed that these neural stem cells derived from the EGFP mice could maintain transgene expression and could differentiate into the MAP2 positive cells after transplantation into the injured spinal cord. The results suggested that NSC expressing EGFP was a useful marker for tracing the cells after transplantation in vivo and functional in the treatment to spinal cord injury.     

Endogenous neural stem cells in central canal of adult rats acquired limited ability to differentiate into neurons following mild spinal cord injury

Endogenous neural stem cells in central canal of adult mammalian spinal cord exhibit stem cell properties following injury. In the present study, the endogenous neural stem cells were labeled with Dil to track the differentiation of cells after mild spinal cord injury (SCI). Compared with 1 and 14 days post mild injury, the number of endogenous neural stem cells significantly increased at the injured site of spinal cord on 3 and 7 days post-injury. Dil-labeled βIII-tublin and GFAP expressing cells could be detected on 7 days post-injury, which indicated that the endogenous neural stem cells in central canal of spinal cord differentiated into different type of neural cells, but there were more differentiated astrocytes than the neurons after injury. Furthermore, after injury the expression of inhibitory Notch1 and Hes1 mRNA began to increase at 6 hours and was evident at 12 and 24 hours, which maintained high levels up to 7 days post-injury. These results indicated that a mild SCI in rat is sufficient to induce endogenous neural stem cells proliferation and differentiation. However, the ability to differentiate into neurons is limited, which may be, at least in part, due to high expression of inhibitory Notch1 and Hes1 genes after injury.     

脐血干细胞移植及电针治疗脊髓损伤大鼠神经生长因子及神经营养因子3的表达

背景：研究表明,脐血干细胞移植对脊髓损伤的恢复起促进作用,而电针也能够通过抑制星形胶质细胞增生,来减少损伤部瘢痕形成,故推测两者结合可能在急性脊髓损伤治疗中发挥重要作用。 目的：观察人脐血干细胞局部移植联合督脉电针治疗后大鼠脊髓损伤组织神经生长因子、神经营养因子3的表达。 方法：选取雌性SD大鼠72只,随机分为对照组、损伤组、移植组、联合组。对照组单纯性背部切口后缝合,损伤组脊髓横断处(T₁₀水平)放置约1 mm×2 mm×2 mm大小、浸润生理盐水的明胶海绵;移植组及联合组在脊髓横断处放置浸润人脐血干细胞悬液的明胶海绵,联合组于造模后1 h开始给予督脉电针治疗。在相应处理7,14,28 d后应用免疫组织化学、Western Blot及实时荧光定量PCR方法检测脊髓组织神经生长因子、神经营养因子3表达量的变化。 结果与结论：脊髓损伤后,移植组与损伤组相比,联合组与移植组相比,神经生长因子、神经营养因子3在7,14,28 d表达量均增加(P < 0.05)。Western Blot、实时荧光定量PCR与免疫组化结果相一致。结果显示人脐血干细胞移植与电针联合治疗脊髓损伤具有协同作用,显著上调损伤脊髓神经生长因子、神经营养因子3的表达水平,有利于脊髓损伤后功能恢复。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

骨髓间充质干细胞静脉移植脊髓损伤大鼠肿瘤坏死因子α及 白细胞介素1β的表达

背景：研究认为间充质干细胞的营养支持在脊髓损伤治疗中起了主要作用,其同损伤宿主神经组织间的相互作用可导致一些不利于损伤修复的炎症因子表达减少。 目的：观察大鼠骨髓间充质干细胞静脉注射移植对脊髓损伤后肿瘤坏死因子α、白细胞介素1β 表达的影响。 方法：运用改良Allen法制备大鼠T10脊髓外伤性截瘫模型,随机分为对照组和骨髓间充质干细胞移植组,设未损伤脊髓的假手术组做对照。骨髓间充质干细胞移植组、假手术组均接受大鼠骨髓间充质干细胞静脉注射移植,对照组静脉注射等量PBS。 结果与结论：对照组和骨髓间充质干细胞移植组损伤脊髓肿瘤坏死因子α、白细胞介素1β蛋白表达较假手术组有明显增加(P < 0.05);骨髓间充质干细胞移植组与对照组比较, 肿瘤坏死因子α、白细胞介素1β蛋白表达受到明显抑制(P < 0.05)。提示大鼠骨髓间充质干细胞静脉移植后能使损伤脊髓局部的肿瘤坏死因子α、白细胞介素1β表达程度降低。这可能是改变脊髓损伤区的微环境,减少脊髓继发性损伤,促进损伤大鼠运动功能恢复的机制之一。