Influence of self‐assembling peptide nanofibre scaffolds on retinal differentiation potential of stem/progenitor cells derived from ciliary pigment epithelial cells

Aim of the study was to investigate the influence of the self‐assembling peptide nanofibre scaffolds (SAPNs) on the growth, proliferation and retinal neuronal differentiation of the stem/progenitor cells (SCs) derived from the ciliary pigment epithelium (CPE) of human cadaveric eye. Here SAPNs (RADA16‐I, PM), which is well described in previous studies, commercially available and xeno‐free. The CPE cells isolated were cultured in DMEM/F12 supplemented with N2 and growth factors such as basic fibroblast growth factor and epidermal growth factor, encapsulated in the scaffolds. The entrapped SCs actively expanded and formed clone‐like clusters in the scaffolds. Many cells in the cluster were proliferating, as revealed by 5‐bromo‐2‐deoxyuridine uptake and could be maintained for up to 6 days and expressed neural progenitor markers such as β‐III tubulin, Nestin, Pax6 and Musashi1. Upon differentiation of these cells in conditioned medium, the cells exhibited retinal neuronal markers such as s‐Opsin, rhodopsin and Recoverin. The RT² profiler polymerase chain reaction array experiments showed selective gene expression, possibly involved in neural stem/progenitor cell adhesion and differentiation. These findings suggest the suitability of the three‐dimensional culture system for the proliferation and maintenance of CPE stem/progenitor cells (CPE‐NS) and for possible use in ex vivo studies of small molecules, drug deliveries for retinal diseases and for use in combination with directed stem/progenitor cell differentiation. and ultimately for tissue replacement therapies. Copyright © 2014 John Wiley & Sons, Ltd.     

Differentiation of human pluripotent stem cells to retinal pigmented epithelium in defined conditions using purified extracellular matrix proteins

A potential application of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) is the generation of retinal pigmented epithelium (RPE) to treat age‐related macular degeneration (AMD), a common but incurable retinal disease. RPE cells derived from hESCs (hESC‐RPEs) and iPSCs (iPSC‐RPEs) express essential RPE markers and can rescue visual function in animal models. However, standard differentiation protocols yield RPE cells at low frequency, especially from iPSC lines, and the common use of Matrigel and xenogeneic feeder cells is not compatible with clinical applications. The extracellular matrix (ECM) can affect differentiation, and therefore changes in ECM composition may improve the frequency of stem cell‐RPE differentiation. We selected several purified ECM proteins and substrates, based on the in vivo RPE ECM environment, and tested their ability to support iPSC‐RPE differentiation and maintenance. iPSCs differentiated on nearly all tested substrates developed pigmented regions, with Matrigel and mouse laminin‐111 supporting the highest pigmentation frequencies. Although iPSC‐RPEs cultured on the majority of the tested substrates expressed key RPE genes, only six substrates supported development of confluent monolayers with normal RPE morphology, including Matrigel and mouse laminin‐111. iPSCs differentiated on mouse laminin‐111 produced iPSC‐RPEs expressing RPE proteins, and hESCs differentiated on mouse laminin‐111 resulted in high yields of functional hESC‐RPEs. This identification of key ECM proteins may assist with future scaffold designs and provide peptide sequences for use in synthetic, xeno‐free, GMP‐compliant generation of RPE from human pluripotent stem cells relevant to clinical translation. Copyright © 2012 John Wiley & Sons, Ltd.     

Embryonic stem cells as a treatment for macular degeneration

Introduction: Retinal degenerations are typically characterized by loss of highly differentiated cell types within the neurosensory retina, such as photoreceptors, or retinal pigment epithelium (RPE). RPE loss is the final common pathway in a number of degenerations including the leading cause of new blindness in the developed world: age-related macular degeneration (AMD).

Areas covered: This paper presents the pathophysiologic case for RPE transplantation with stem cell (SC)-derived tissue, a review of the preclinical data substantiating the hypothesis and the initial clinical trials safety data from early human trials.

Expert opinion: Targeting the RPE for transplantation with SC-derived tissue presents a reasonable therapeutic opportunity in a variety of important, otherwise untreatable, blinding conditions. Success of cellular replacement strategies is contingent on finding a viable source of replacement cells, establishing a safe technique for delivery and survival of transplanted cells within the host, restoration of normal retinal architecture and stabilization or improvement of vision.     

改善微囊化人视网膜色素上皮细胞生长状态的研究

目的 观察上皮细胞培养液中添加果糖、成纤维细胞生长因子和牛磺酸后对人视网膜色素上皮(hRPE)细胞海藻酸钠-壳聚糖-海藻酸钠微囊化后的细胞总数、存活率及活细胞数的影响.方法 将hRPE细胞微囊化后将其分为4组,一组作为空白对照组(空白组)只加入上皮细胞培养液、另外3组在培养液中分别加入果糖(果糖组)、成纤维细胞生长因子(成纤维细胞生长因子组)、牛磺酸(牛磺酸组)后培养7 d,在0、1、3、7 d测定微囊内细胞的细胞总数、存活率及活细胞数.结果 4组细胞微囊化后的hRPE细胞至培养7 d,细胞存活率最低为(75.00±3.00)%,但各组间差异无统计学意义(P均＞0.05);而微囊内细胞总数牛磺酸组(7.20±0.36)×104、成纤维细胞生长因子组(8.00±0.46)×104,比空白对照组(6.10±0.56)×104明显增多(t值分别为-2.872、-4.564,P均＜0.05),果糖组(6.00±0.46)×104与空白组比较差异无统计学意义(P＞0.05).结论 细胞生长因子、牛磺酸可促进海藻酸钠-壳聚糖-海藻酸钠微囊内的hRPE细胞的增殖.

Abstract：

Objective To observe the effects of fructose,fibroblast growth supplement(FGS) and ethylamine sulfonic acid on the total number,the survival rate and the survival number of Human Retinal Pigment Epithelial(hRPE) Cell.Methods Microencapsulated hRPE cells were plated and cultured in four kinds of mediums,which contained fructose,fibroblast growth supplement(FGS),ethylamine sulfonic acid or no extra ingredient respectively.The total cell number,survival rate and viable cell number of the microencapsulated hRPE cell on day 0th,1st,3rd,7th were calculated.Results After 7days of culture,the lowest cell survival rate of microencapsulated hRPE cells in the four groups was(75.00±3.00)%,but there were no significantly differences(Ps＞0.05) among the groups.The total number of cells in the fibroblast growth factor group([8.00±0.46]×104) and ethylamine sulfonic acid group([7.20±0.36]×104) were significantly higher than the blank group(([6.10±0.56]×104),Ps＜0.05),while no statistical difference was observed in the comparison between the fructose group([6.00±0.46]×104) and blank control(P＞0.05).Conclusion The FGS and ethylamine sulfonic acid can promote the proliferation of the microencapsulated hRPE cells.     

Vitrification and storage of oral mucosa epithelial cell sheets

Shipping time and shipping delays might affect the quality of the stem cells based engineered “organs.” In our laboratory, we have developed a limbal stem cell deficient (LSCD) rabbit model. To reverse the LSCD, we cultured oral mucosal epithelial cells for 2–3 weeks and engineered cultured autologous oral mucosa epithelial cell sheets (CAOMECS), which were grafted on the LSCD cornea. The purpose of this study was to vitrify CAOMECS and to store it until the CAOMECS can be grafted onto patients. CAOMECS were vitrified in LN₂ for up to 204 days. We tested two different methods of vitrification with different solutions; however, CAOMECS were only viable when they were not stored in a vitrification solution; results were only reported from this CAOMECS. On the basis of hematoxylin and eosin staining, we showed that the CAOMECS morphology was well preserved after long‐term storage in LN₂. Most of the preservation solutions maintained the CAOMECS phenotype (Ki67, proliferating cell nuclear antigen (PCNA), Beta‐Catenin, ZO‐1, E‐Cadherin, CK3, CK4, CK13). The exception was the solution composed with ethylene glycol and Dimethyl sulfoxide (DMSO): this resulted in loss of DeltaN‐p63 expression. DeltaN‐p63 is an important marker for cell proliferation. The expression of proteins involved in cell–cell connection and the differentiation markers were maintained. Apoptosis was not detected in the thawed CAOMECS. We demonstrated that CAOMECS can be stored long‐term in LN₂ without affecting their morphology and phenotype.     

Collagen vitrigels with low‐fibril density enhance human embryonic stem cell‐derived retinal pigment epithelial cell maturation

Structural and biochemical cues of extracellular matrix can substantially influence the differentiation and maturation of cultured retinal pigment epithelial (RPE) cells. In this study, thin collagen vitrigels were engineered to create collagen nanofibrillar structures of different fibril densities in an effort to evaluate the maturation of human embryonic stem cell–derived retinal pigment epithelial (hESC‐RPE) cells. The ultrastructure of the different collagen vitrigels was characterized by transmission electron microscopy, and the mechanical properties were evaluated by tensile testing. The pigmentation and polarization of cells, in addition to key RPE marker gene and protein expression levels, were analyzed to determine the differentiation of hESCs on the gels. The hESC‐RPE differentiation was most significant in collagen vitrigels with low fibril density with mature collagen fibrils with diameter of around 60 nm and Young's modulus of 2.41 ± 0.13 MPa. This study provides insight into the influence of collagen nanofibrillar structures on hESC‐RPE maturation and presents a potential bioengineered substratum for hESC‐RPE for future preclinical and clinical applications.     

A simple and static preservation system for shipping retinal pigment epithelium cell sheets

The ability to move cells and tissues from bench to bedside is an essential aspect of regenerative medicine. In this study, we propose a simple and static shipping system to deliver tissue‐engineered cell sheets. Notably, this system is electronic‐device‐free and simplified to minimize the number of packing and opening steps involved. Shipping conditions were optimized, and application and verification of the system were performed using human iPS cell‐derived or fetal retinal pigment epithelium (RPE) cell sheets. The temperature of the compartments within the insulated container was stable at various conditions, and filling up the cell vessel with medium effectively prevented turbulence‐induced mechanical damage to the RPE cell sheets. Furthermore, no abnormal changes were observed in RPE morphology, transepithelial electrical resistance, or mRNA expression after transit by train and car. Taken together, our simple shipping system has the potential to minimize the costs and human error associated with bench to bedside tissue transfer. This specially designed regenerative tissue shipping system, validated for use in this field, can be used without any special training. This study provides a procedure for easily sharing engineered tissues with the goal of promoting collaboration between laboratories and hospitals and enhancing patient care.     

Intraperitoneal administration of adipose tissue‐derived stem cells for the rescue of retinal degeneration in a mouse model via indigenous CNTF up‐regulation by IL‐6

As the world's population begins to age, retinal degeneration is an increasing problem, and various treatment modalities are being developed. However, there have been no therapies for degenerative retinal conditions that are not characterized by neovascularization. We investigated whether transplantation of mouse adipose tissue‐derived stem cells (mADSC) into the intraperitoneal space has a rescue effect on NaIO₃‐induced retinal degeneration in mice. In this study, mADSC transplantation recovered visual function and preserved the retinal outer layer structure compared to the control group without any integration of mADSC into the retina. Moreover, endogenous ciliary neurotrophic factor (CNTF) was elevated in the retinas of mADSC‐treated mice. We found that lipopolysaccharide (LPS) or LPS‐stimulated monocyte supernatant induced the secretion of granulocyte colony stimulating factor (GCSF), CD54, CXCL10, interleukin‐6 (IL‐6), and CCL5 from the mADSC by cytokine array. Network inference was conducted to investigate signaling networks related to CNTF regulation. Based on bioinformatics data, the expression of IL‐6 was related to the expression of CNTF. Additionally, intravitreal injection of IL‐6 in rats produced up‐regulation of endogenous CNTF in the retina. mADSC had a rescue effect on retinal degeneration through the up‐regulation of endogenous CNTF by IL‐6. Thus, transplantation of mADSC could be a potential treatment option for retinal degeneration.     

丹红注射液联合骨髓间充质干细胞移植治疗大鼠脑梗死

背景：单纯骨髓间充质干细胞移植修复受损脑组织的作用并不十分理想。目的：观察丹红注射液联合骨髓间充质干细胞移植治疗大鼠脑梗死的效果。方法：用线栓法制备大鼠大脑中动脉阻塞模型,随机分为3组,模型组尾静脉注射PBS、丹红注射液组尾静脉注射2mL／kg丹红注射液、联合治疗组联合注射2mL／kg丹红注射液＋2．0×10^9L^-1的骨髓间充质干细胞悬液,连续5d,1次／d。结果与结论：在骨髓间充质干细胞移植后2周,联合治疗组神经功能评分明显优于模型组及丹红注射液组（P〈0．05）;移植后3周联合治疗组大鼠脑梗死体积明显小于模型组和丹红注射液组（P〈0．05）;病理组织学观察也可见联合治疗组的组织损伤减轻程度大于丹红注射液组和模型组。结果可见丹红注射液联合骨髓间充质干细胞移植治疗大鼠脑梗死疗效显著,可以对脑细胞起到保护作用。     

视网膜色素上皮细胞对结膜囊成纤维细胞的影响

目的 :观察人视网膜色素上皮 (RPE)细胞调理液 (RPE -CM )对人结膜囊成纤维细胞增殖及胶原合成的影响。方法 :用MTT比色法、3H -脯氨酸掺入法和原位杂交法观察RPE -CM对体外培养的成纤维细胞生长及胶原合成活性的作用。结果 :RPE细胞培养调理液可使成纤维细胞的OD值明显增高 (P<0.01) ,还可使成纤维细胞3H -脯氨酸掺入的CPM值明显高于对照组 (P<0.01)。处理组成纤维细胞中Ⅰ型前胶原mRNA原位杂交信号明显增强。结论 :RPE -CM可刺激成纤维细胞生长及胶原合成 ,此作用与RPE细胞的去分化程度有关     

A bilayer photoreceptor‐retinal tissue model with gradient cell density design: A study of microvalve‐based bioprinting

ARPE‐19 and Y79 cells were precisely and effectively delivered to form an in vitro retinal tissue model via 3D cell bioprinting technology. The samples were characterized by cell viability assay, haematoxylin and eosin and immunofluorescent staining, scanning electrical microscopy and confocal microscopy, and so forth. The bioprinted ARPE‐19 cells formed a high‐quality cell monolayer in 14 days. Manually seeded ARPE‐19 cells were poorly controlled during and after cell seeding, and they aggregated to form uneven cell layer. The Y79 cells were subsequently bioprinted on the ARPE‐19 cell monolayer to form 2 distinctive patterns. The microvalve‐based bioprinting is efficient and accurate to build the in vitro tissue models with the potential to provide similar pathological responses and mechanism to human diseases, to mimic the phenotypic endpoints that are comparable with clinical studies, and to provide a realistic prediction of clinical efficacy.     

胚胎干细胞移植治疗脑血管疾病

背景：胚胎干细胞移植是否能够成为脑血管疾病治疗有效的方法已成为目前研究的热点。目的：探讨胚胎干细胞分化神经前体细胞移植治疗脑血管疾病的效果及可行性。方法：分别建立帕金森病、缺血性脑损伤以及血管性痴呆大鼠模型,并进行胚胎干细胞体外培养,诱导分化为神经前体细胞,将胚胎干细胞分化神经前体细胞移植入相应脑血管疾病模型大鼠脑内,观察脑血管病变大鼠的旋转行为学变化、脑组织病理变化以及海马结构的变化和神经细胞数目的变化。结果与结论：胚胎干细胞分化神经前体细胞移植入帕金森病大鼠脑内后,阿朴吗啡诱发的旋转次数逐渐减少,呈下降趋势,纹状体多巴胺的含量明显增高。胚胎干细胞分化神经前体细胞移植入缺血性脑损伤大鼠脑内后,能够长期存在于脑内,并迁移、分布于受损的海马,构成海马结构,进一步分化为神经元,并且受损海马内的神经细胞数量明显增加。说明移植的胶质细胞源性神经营养因子基因修饰的胚胎干细胞可改善血管性痴呆大鼠的学习记忆功能,增强神经的可塑性,诱导自身定向迁移并分化为成熟神经元。     

亲缘异基因造血干细胞移植联合甲磺酸伊马替尼治疗Ph阳性白血病

背景：造血干细胞移植是可以治愈Ph＋白血病有效方法,甲磺酸伊马替尼是一种高度特异的酪氨酸激酶抑制剂,能抑制BCR/ABL酪氨酸激酶活性,在Ph＋白血病中的应用越来越多。目的：探讨甲磺酸伊马替尼联合亲缘异基因造血干细胞移植治疗Ph＋白血病的临床疗效。方法：回顾性分析2011年1月至2012年10月采用亲缘异基因造血干细胞移植联合甲磺酸伊马替尼治疗12例Ph＋白血病的疗效并文献复习。结果与结论：12例患者移植后均获得造血重建,移植后中性粒细胞和血小板植活的中位时间分别为15 d和18 d;发生Ⅱ度急性移植物抗宿主病7例,Ⅲ度急性移植物抗宿主病1例,局限型慢性移植物抗宿主病7例,广泛型慢性移植物抗宿主病3例;无白血病存活率为67%,移植相关死亡率为25%。行HLA匹配亲缘造血干细胞移植者的总体存活率为75.0%。平均无病生存8.5个月（7-17个月）,BCR/ABL转阴时间2-5个月。亲缘异基因造血干细胞移植前、后联合甲磺酸伊马替尼治疗Ph＋白血病,具有降低移植前白血病细胞负荷,抑制残留白血病细胞增殖,促进供者完全嵌合状态的转变,是一种安全有效的治疗方法。     

自体外周血干细胞移植治疗周围神经损伤

背景：关于神经干细胞对周围神经损伤的治疗已有多篇报道,但外周血干细胞对周围神经损伤治疗鲜有报道。目的：探讨自体外周血干细胞移植治疗周围神经损伤使失神经骨骼肌重获神经再支配的临床应用。方法：应用外周血干细胞治疗周围神经损伤6例,同时与周围神经损伤单纯行神经断端吻合或神经移植10例比较。2组患者术后常规肌注鼠神经生长因子一两个疗程,同时给予针灸、理疗、经皮电刺激治疗及功能康复训练。结果与结论：两组患者随访均超过6个月。干细胞移植组运动神经传导速度和感觉神经传导速度的恢复率要明显高于单纯神经吻合组。提示周围神经损伤后给予修复局部用外周血干细胞移植能够使远端失神经骨骼肌早期重新获得神经再支配。     

Effects of cell concentration,time of fresh storage,and cryopreservation on peripheral blood stem cells: PBSC fresh storage and cryopreservation

IntroductionPeripheral blood stem cells are widely used in autologous or allogeneic transplantation. The quality of the product directly impacts clinical outcomes, and the cell quality and/or functionality may be influenced by the storage conditions as time, temperature, total nucleated cells (TNC) concentration and cryopreservation requirement.ObjectiveTo verify the effects of time, cell concentration, and cryopreservation/thawing in the viability and functionality of stem cells for transplantation.MethodsWe evaluated TNC, CD45⁺ viable cells, CD34⁺ viable cells, and cell viability and functionality of 11 samples. Measurements were performed immediately and 24 h, 48 h, 72 h, and 96 h after sample collection at high and low TNC concentrations. The same parameters were also evaluated after cryopreservation and thawing of the samples.ResultDuration of storage and TNC concentration exhibited a negative effect on cell quality (CD45⁺ viable cells, CD34⁺ viable cells and functionality). Moreover, the association of these parameters increased the negative effect on graft quality. Cryopreservation and thawing also negatively affected the collected sample regarding viable CD34⁺ cells (recovery 66.2 %), viable CD45⁺ cells (recovery 56.8 %), and 7-AAD viability. No significant losses in viable CD45⁺/CD34⁺ cells and functionality were observed in the first 24 h in both TNC conditions.ConclusionThese results emphasize the importance to consider carefully the storage conditions until transplantation, measuring TNC/μL until 24 h after collection (diluting the product when TNC > 300 × 10³/μL) and infusing fresh graft as soon as possible.     

猕猴单倍体相合非清髓造血干细胞联合间充质干细胞移植的实验研究

目的 用非清髓预处理建立猕猴单倍体相合造血干细胞移植(HSCT)模型，研究间充质干细胞(MSC)在单倍体相合移植中的作用。方法采用健康、单倍体相合的猕猴亲子配对，受非清髓性预处理用氟达拉滨 环磷酰胺 ^60Co(200cGy)全身照射 兔抗人胸腺细胞球蛋白，移植物抗宿主病(GVHD)预防用环孢菌素A、霉酚酸酯、抗CD25单抗。实验分为单纯造血干细胞(HSC)移植组和HSC联合MSC移植组；检测供受体嵌合水平，观察造血恢复、GVHD等情况。结果单倍体相合的子代猕猴采用非清髓预处理方案，可获稳定植入?比较了单纯非清髓HSC移植组和HSC联合MSC移植组的造血恢复，发现造血恢复时间主要与嵌合状态有关；MSC可促进植入；相同条件下，HSC联合MSC移植组更容易形成供受混合嵌合；GVHD的发生率低。结论成功建立了猕猴非清髓性单倍体相合HSCT的模型，非清髓HSCT联合MSC可能在单倍体相合移植中有较好效果。     

Clinically applicable transplantation procedure of dermal papilla cells for hair follicle regeneration

Dermal papilla cells (DPCs) interact with epithelial stem cells and induce hair folliculogenesis. Cell-based therapies using expanded DPCs for hair regeneration have been unsuccessful in humans. Two major challenges remain: first, expanded DPCs obtained from adult hair follicles have functional limitations; second, a clinically applicable method is needed for transplanting DPCs. This study aimed to identify an efficient, minimally invasive and economical DPC transplantation procedure for use in clinical settings. Five clinically applicable transplantation procedures were tested, termed the Pinhole, Laser, Slit, Non-vascularized sandwich (NVS) and Hemi-vascularized sandwich (HVS) methods. Labelled rat dermal papilla tissue was transplanted into rat sole skin, and hair follicle regeneration was evaluated histologically. Regenerated follicles and labelled DPCs were detected for all methods, although some follicles showed abnormal growth, i.e. a cystic or inverted appearance. The HVS method, pioneered here, resulted in significantly larger number of regenerated follicles that were more mature and regular than those observed using the other methods. Moreover, hair growth was detected after expanded adult-derived DPC transplantation using the HVS method. These results suggest that direct contact of epithelial and dermal components and better vascularization/oxygenation of the recipient site are critical for hair regeneration in cell-based therapies.     

依达拉奉联合神经干细胞移植修复大鼠脊髓损伤

背景：依达拉奉是一种自由基清除药,可以减轻受损神经组织水肿和改善脊髓损伤区微环境。目的：观察依达拉奉联合神经干细胞移植对大鼠脊髓全横断损伤的修复效果。方法：成年雌性SD大鼠80只,建立胸9脊髓全横断损伤模型,随机分为4组：对照组不做处理;依达拉奉组脊髓损伤后6h经尾静脉注射依达拉奉;神经干细胞移植组脊髓损伤后6h脊髓损伤区域注入神经干细胞悬液;依达拉奉＋细胞移植组脊髓损伤后6h神经干细胞移植的同时尾静脉注射依达拉奉。结果与结论：造模后8周可观察到PKH-26标记的神经干细胞在体内存活并在脊髓内迁移;细胞移植组和依达拉奉联＋细胞移植组可见少量连续性神经纤维通过损伤区。荧光金逆行脊髓追踪显示神经干细胞移植组和依达拉奉＋细胞移植组可见被荧光金标记的神经锥体细胞穿越损伤区。PKH-26标记的阳性细胞数及荧光金阳性神经纤维数：依达拉奉＋细胞移植组最多,依达拉奉组、神经干细胞移植组次之,对照组最少,各组之间差异有显著性意义（P〈0.05）;后肢功能运动BBB评分依次为依达拉奉＋细胞移植组〉神经干细胞移植组〉依达拉奉组〉对照组。提示依达拉奉能促进神经干细胞在损伤区的存活并向神经细胞分化,依达拉奉联合神经干细胞移植有促进细胞移植修复大鼠脊髓损伤的效果。     

球后注射脐血干细胞治疗老年性黄斑变性的护理

目的:探讨球后注射脐血干细胞结合针刺治疗老年性黄斑变性的护理方法及作用。方法:对我院9例采用球后注射脐血干细胞治疗的患者的护理方法进行归纳总结。结果:所有患者术程顺利,未出现任何并发症。结论:术前、术中、术后分期护理,以及心理护理,可稳定患者的心理,提高患者配合度,降低并发症的发生。     

自体骨髓间充质干细胞移植治疗脑梗死的临床分析

目的：探讨自体骨髓间充质干细胞经蛛网膜下腔移植治疗脑梗死的临床疗效及安全性。方法选择2011年1月至2012年7月在江西省人民医院神经科住院的60例脑梗死患者,随机分为治疗组和对照组,每组各30例。对照组给予常规药物治疗,并尽早进行康复训练;治疗组在对照组治疗的基础上,行自体骨髓间充质干细胞经蛛网膜下腔移植治疗,每周1次,两周为1个疗程。分别于移植前,移植后3、6个月对两组患者进行神经功能缺损程度评定（NIHSS评分）和日常生活活动量表（Barthel指数）评分,同时观察治疗组患者的不良反应。结果两组患者移植前NIHSS评分及Barthel指数评分与对照组比较,差异无统计学意义（P＞0．05）;治疗组移植后3、6个月的NIHSS评分均明显低于对照组（P＜0．01）,而Barthel指数评分明显高于对照组（P＜0．01）。治疗组移植术后3例出现低热,4例出现头痛,均未经治疗症状自行消失。结论自体骨髓间质干细胞经蛛网膜下腔移植治疗脑梗死是安全可行的,可明显改善脑缺血后神经功能损伤,为脑梗死的治疗开辟了一条新的途径。