人羊膜上皮细胞移植改善AD样病变模型大鼠学习记忆能力

目的:观察人羊膜上皮细胞(h AECs)对阿尔茨海默病(AD)样病变大鼠模型的治疗效应。方法:采用胰蛋白酶消化法分离h AECs,流式细胞术分析表型。48只雄性SD大鼠随机分为假手术组、模型组、培养基组和h AECs移植组,每组12只。采用双侧脑室注入脂多糖(LPS)复制AD样病变大鼠模型。AD样病变大鼠海马区移植5×105个h AECs。细胞移植后2周,Morris水迷宫试验观察行为学变化,HE和硫磺素S染色观察海马病理变化,免疫组化染色检测β-淀粉样蛋白42(Aβ42)、Tau蛋白和乙酰胆碱(ACh)的变化,流式细胞术检测外周血T淋巴细胞亚群的变化,流式微球阵列捕获技术(cytometric bead array,CBA)检测血清细胞因子含量,免疫荧光染色检测海马区人细胞核抗原阳性细胞及其神经元特异性核蛋白(Neu N)的表达。结果:与模型组和培养基组比较,h AECs移植组大鼠逃避潜伏期明显缩短(P0.01),跨域平台次数明显增加(P0.05);海马神经元病损减轻,Aβ沉积减轻(P0.05),磷酸化Tau蛋白水平下降(P0.05),ACh增加(P0.05);外周血Th1和Th17细胞百分比下降(P0.05),而Th2和Treg细胞升高(P0.05);IL-2和IFN-γ水平下调(P0.05),而IL-4上调(P0.05);移植区可见h AECs,并表达Neu N。结论:h AECs可明显改善AD样病变模型大鼠空间辨别性学习记忆能力,减轻海马病理损伤,其免疫调节效应可能发挥重要作用。     

立体定向移植骨髓间充质干细胞改善血管性大鼠的学习记忆能力

背景：大量的实验证明将骨髓间充质干细胞移植到复制的中枢神经系统疾病大鼠模型后,能迁移到损伤部位,表达神经细胞的潜在特性并且提高神经功能。 目的：验证骨髓间充质干细胞对拟人类血管性痴呆模型大鼠学习记忆障碍的改善作用。 方法：分离SD大鼠骨髓间充质干细胞,于细胞移植前掺入10 mg/L的BrdU进行标记。建立Wistar大鼠血管性痴呆样学习记忆障碍动物模型,随机分为模型组、假手术组与移植组。移植组致伤后第14天,通过立体定向途径移植骨髓间充质干细胞到大鼠海马,假手术组给予等量生理盐水,模型组大鼠不做处理。采用Morris水迷宫检测大鼠空间学习记忆能力。损伤后第90天处死大鼠,观察海马组织有无Brdu+神经元特异性烯醇化酶、Brdu+胶质纤维酸性蛋白免疫组织化学双染阳性细胞,并且观察从侧脑室到海马是否存在神经元前体细胞的标志Doublecortin (DCX)吻侧迁移流。 结果与结论：①移植组大鼠采用Morris水迷宫检测大鼠空间学习记忆能力逃避潜伏期及跨越平台次数优于模型组及假手术组(P < 0.05)。②移植组大鼠海马组织及其周围可见免疫组织化学双染阳性细胞,但未见从侧脑室到海马关于神经元前体细胞的标志Doublecortin (DCX)吻侧迁移流。结果提示,骨髓间充质干细胞移植可以促进脑损伤大鼠的神经功能的恢复,其机制可能与移植细胞分化为神经元样和神经胶质细胞样细胞,并分泌或促进宿主分泌神经营养因子有关。     

阿尔茨海默病与Wnt信号通路及神经干细胞的关系

背景：阿尔茨海默病的病因涉及遗传、环境、免疫等多种因素和机制,大量研究表明Wnt信号通路与之密切相关,也有研究表明,Wnt信号通路对神经干细胞的增殖发挥着决定性作用。 目的：对阿尔茨海默病的病理过程与Wnt信号通路的关系以及Wnt信号通路与神经干细胞增殖分化进行综述,为阿尔茨海默病的治疗提供理论依据。 方法：通过Pubmed数据库检索有关阿尔茨海默病病理过程与Wnt信号通路及Wnt信号通路与神经干细胞关联的相关文献,检索词为“Alzheimer’s disease、Wnt signaling pathway、NSCs、stem cell differentiation、amyloid-β protein、Protein Tau”。纳入与阿尔茨海默病和Wnt信号通路相关的文献,排除重复性研究,保留50篇文献进行综述。 结果与结论：目前通过神经干细胞移植来治疗以神经元的缺失为特征的神经退行性疾病已成为研究的热点,而如何调控神经干细胞向特定神经元分化成为了研究的难点,信号转导在神经干细胞的分化中起重要的作用,其中Wnt信号通路是调节神经干细胞增殖及分化的细胞外的重要因素。Wnt信号通路的失活可以促进阿尔茨海默病的病理过程,相反激活Wnt信号通路可以保护海马神经元,同时促进神经干细胞的分化,为阿尔茨海默病的治疗提供新的思路。     

石菖蒲不同药效部位改善阿尔茨海默病模型小鼠的认知功能

目的: 探讨石菖蒲不同药效部位改善阿尔茨海默病(Alzheimer disease, AD)模型小鼠认知功能的机制。方法: 雄性NIH小鼠,采用双侧海马CA1内注射淀粉样β蛋白(Aβ_1-42,2 g/L,每侧2 μL)制备AD小鼠动物模型,并以生理盐水模拟注射和不注射的正常组小鼠为对照。通过水迷宫测试,筛选出与两对照组均有显著差异的认知障碍小鼠,留作AD动物模型。AD动物模型又随机分为生理盐水灌胃组、石菖蒲水煎液灌胃组、石菖蒲去油水煎液灌胃组和石菖蒲挥发油灌胃组。每组6只,共4组。以生理盐水灌胃组为对照,石菖蒲不同药效部位灌胃组每天对应灌胃1次(0.2 g 石菖蒲/10 g体重),连续3周。灌胃结束,Morris水迷宫检测小鼠的空间学习记忆能力,测定小鼠大脑和海马中一氧化氮合酶(nitric oxide synthase, NOS)活性,免疫组织化学方法检测脑中NOS的表达。结果: 水迷宫测试结果表明,石菖蒲不同药效部位(水煎液和挥发油)灌胃2组模型小鼠与生理盐水灌胃组小鼠相比,60 s内跨越平台的次数和在目标象限的探索时间均明显增加(P<0.05);大脑和海马内NOS活性检测显示,石菖蒲不同药效部位灌胃3组AD模型小鼠和其对照组相比,大脑和海马内NOS活性下降,NOS阳性神经元数量明显减少(P<0.05)。结论: 石菖蒲不同药效部位能显著改善Aβ_1-42致AD模型小鼠的学习记忆能力,其疗效可能与降低大脑和海马中的NOS活性有关。     

吡格列酮联合骨髓间充质干细胞移植治疗改善大鼠心肌梗死后的心功能

背景：前期研究发现骨髓间充质干细胞移植能够改善心肌梗死大鼠的心功能,但整体效果并不太理想。 目的：拟采用PPAR-γ激动剂吡格列酮联合骨髓间充质干细胞移植治疗以进一步改善心肌梗死大鼠的心功能并探讨相关机制。 方法：开胸结扎20只SD大鼠左前降支冠状动脉并随机分为2组：骨髓间充质干细胞组和骨髓间充质干细胞+吡格列酮组。2周后在局部梗死心肌内注射PKH26标记的由PBS悬浮的骨髓间充质干细胞,联合治疗组在注射骨髓间充质干细胞后予以吡格列酮3 mg/(kg•d)连续灌胃2周。细胞移植后2周进行超声心动图检测,免疫荧光染色、Western blot、qRT-PCR检测左心室心肌组织不同区域PPAR-γ、TGF-β1/SMAD通路相关因子和Cx43的表达情况。 结果与结论：两组大鼠基础心功能参数无明显差异性。细胞移植2周后,骨髓间充质干细胞+吡格列酮组左室舒张末径、左室收缩末径明显减小,左室射血分数明显增高;左心室心肌组织不同区域PPAR-γ和Cx43的表达量显著增加;TGF-β1、SMAD2、SMAD3在梗死区和梗死边缘区表达明显下降。以上结果提示PPAR-γ激动剂吡格列酮干预能够增强骨髓间充质干细胞移植对心功能的改善作用,其机制可能与PPAR-γ抑制TGF-β1/SMAD通路进而提高Cx43的表达有关。  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     

中脑神经干细胞和骨髓基质干细胞对帕金森大鼠行为学及脑组织形态学的影响

背景：研究发现神经干细胞或者骨髓基质干细胞可脑内移植治疗中枢神经系统疾病,但是两种干细胞移植治疗效果的比较相对较少。目的：观察比较中脑神经干细胞和骨髓基质干细胞对帕金森大鼠行为学及脑组织形态学的影响。方法：58只SD大鼠构建帕金森病模型,随机分为3组,分别为骨髓基质干细胞组(20只)、中脑神经干细胞组(20只)、生理盐水组(18只)。造模成功后3周,选取右侧纹状体 2 个坐标点注入骨髓基质干细胞悬液、中脑神经干细胞悬液、生理盐水5 μL。移植后5个月腹腔注射阿朴吗啡观察大鼠的行为学变化,取纹状体脑组织制备石蜡切片进行免疫组化荧光染色。结果与结论：治疗后第5个月骨髓基质干细胞组和中脑神经干细胞组大鼠旋转次数较治疗前显著下降(P < 0.05),且明显低于生理盐水组(P < 0.05),骨髓基质干细胞组和神经干细胞组比较差异无显著性意义(P > 0.05)。移植后第1周,骨髓基质干细胞组纹状体内有BrdU与Nestin双重染色细胞;移植后第1个月,脑纹状体内出现BrdU/GFAP双重染色细胞和BrdU/NSE双重染色细胞;纹状体内存在TH阳性细胞,但未出现明显BrdU/TH双重染色细胞;移植1个月之后,BrdU与Nestin双重染色细胞数量逐渐减少,最终基本消失,但仍然存在一定的数量的BrdU/GFAP、BrdU/NSE双重染色细胞,BrdU/GFAP阳性细胞数量相对较多。同一时间点,中脑神经干细胞组也存在类似情况,但生理盐水组未发现双标细胞。结果表明,中脑神经干细胞和骨髓基质干细胞移植均可改善帕金森大鼠的行为学,且可分化为神经元、星形胶质细胞、多巴胺能神经元。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     

静脉移植骨髓基质干细胞对脑缺血大鼠神经功能及神经细胞凋亡的影响

目的： 探讨骨髓基质干细胞静脉移植后进入大鼠局灶性缺血脑内的短期存活情况及其对脑缺血大鼠神经功能的影响。方法: 将BrdU标记大鼠骨髓基质干细胞后，经尾静脉注射到局灶性脑缺血大鼠体内，分别在脑缺血术后1 d、7 d、14 d和28 d通过神经功能评分观察移植后大鼠神经行为学变化情况，通过组织学方法观察移植到脑内的骨髓基质干细胞存活状态和脑内缺血区与非缺血区神经细胞死亡情况，用RT-PCR方法观察到骨髓基质干细胞表达细胞因子及神经营养因子。结果: 骨髓基质干细胞移植组在14 d和28 d大鼠神经功能评分显著低于对照组（P<0.05），神经功能得到明显改善。静脉移植的骨髓基质干细胞在移植的1 d主要分布在损伤侧大脑中动脉周围组织间质中，在第3d沿着损伤侧的下丘脑迁移至海马的CA1(cornu ammonis 1, CA1)区；骨髓基质干细胞移植组大鼠梗死灶周围的死亡细胞在14 d和28 d明显少于对照组（P<0.05）。结论: 经静脉注射移植骨髓基质干细胞能迁移到损伤区并能够明显促进局灶性脑缺血大鼠的神经行为功能恢复；抗凋亡、分泌神经营养因子改善微环境、动员神经干细胞并迁移至缺血区可能是静脉注射骨髓基质干细胞治疗局灶性脑缺血的机制。     

雌激素对阿尔茨海默病去势雌性SD大鼠学习记忆能力的影响

目的观察雌激素对阿尔茨海默病(AD)大鼠学习记忆能力的影响。方法首先采用Aβ1～40,1μL(10μg/μL)立体定位SD大鼠单侧海马内注射建立AD动物模型,二周后双侧卵巢切除术(OVX)制备去卵巢大鼠模型后给予雌激素替代治疗(ERT),最后通过Morris水迷宫观察动物模型的学习、记忆能力变化情况。结果 ERT组AD动物模型的水迷宫逃避潜伏期和目标象限游泳时间比OVX组明显缩短(P0.05)。结论雌激素具有改善AD大鼠模型认知功能的作用。     

外源性硫化氢对阿尔茨海默病大鼠学习能力及海马形态学的影响

目的:观察外源性硫化氢对阿尔茨海默病大鼠的空间学习记忆功能及海马组织形态学的影响。方法:通过双侧海马内注射Aβ25-35制作大鼠AD模型,再连续7 d脑室内给予硫氢化钠。采用Morris水迷宫测试大鼠认知功能,断头取脑行HE染色、透射电镜等方法观察海马神经元的结构变化。结果:海马内注射Aβ25-35后大鼠学习记忆能力明显下降,可见海马细胞排列紊乱,核边聚、碎裂。脑室内给予硫氢化钠可显著改善大鼠认知功能,减轻海马神经元病变。结论:外源性硫化氢对Aβ25-35诱导的阿尔茨海默病模型大鼠脑组织内海马退行性变神经元有保护作用,并能改善大鼠学习记忆能力。     

不同途径移植骨髓间充质干细胞改善大鼠肝硬化

背景：骨髓间充质干细胞移植可减轻肝硬化程度,改善肝功能。 目的：观察不同途径移植骨髓间充质干细胞对四氯化碳诱导大鼠肝硬化的作用。 方法：将60只SD大鼠随机分为正常组、对照组、门静脉移植组、肝动脉移植组、尾静脉移植组,后4组采用四氯化碳联合乙醇制作肝硬化模型,对照组不进行移植,其余3组分别经门静脉、肝动脉、尾静脉移植大鼠骨髓间充质干细胞1×10⁶。 结果与结论：移植4周后,与对照组比较,移植3组大鼠肝功能均得到明显改善,血清白蛋白、胆碱酯酶显著升高(P < 0.05),转氨酶、胆红素、凝血时间、Ⅳ型胶原显著降低(P < 0.05),肝纤维化程度显著减轻(P < 0.05)。门静脉移植组及肝动脉移植组优于尾静脉移植组,前两者之间差异无显著性意义(P > 0.05)。说明经门静脉、肝动脉、尾静脉移植骨髓间充质干细胞均可减轻肝纤维化程度,改善肝功能,但肝动脉及门静脉移植途径优于外周血静脉途径。     

神经干细胞移植治疗老年痴呆

BACKGROUND:Neural stem cell transplantation has been used to treat a series of brain injury diseases, such as cerebral palsy, but its effect on Alzheimer’s disease is rarely reported. OBJECTIVE:To observe the effect of neural stem cell transplantation on the behavior and immune regulating system of Alzheimer’s disease rats.  METHODS:Thirty-five Sprague-Dawley rats were enrolled to make a postcerebral incision and given hippocampal injection of amanita phalloides acid to establish rat models of Alzheimer’s disease. Another 10 rats were only given hippocampal injection of normal saline after preparation of postcerebral skin incision as sham operation group. Then 32 successful rat models were randomly divided into two groups (n=16 per group): rats in experimental group were administrated hippocamal injection of 5×109/L allogeneic neural stem cell suspension; those in model group were given no injection. Five-day Morris water maze test was conducted at 4 weeks after transplantation. At 1 week after Morris water maze test, levels of interleukin-1 and interleukin-10 in the cerebral homogenate were detected, as well as pathological changes of brain tissues were observed in the three groups. RESULTS AND CONCLUSION:Compared with the model group, the abilities of cognition and memory were significantly higher in the sham operation group (P < 0.01), and the abilities of spatial learning and memory were significantly higher in the experimental group (P < 0.05, P < 0.01). Levels of interleukin-1 and interleukin-10 in the model group were significantly higher than those in the sham operation group (P < 0.01) but significantly lower than those in the experimental group (P < 0.01). Besides, the number of neurons in the model group was obviously less than that in the experimental and sham operation group. These results indicate that neural stem cell transplantation supplements and protects neurons against Alzheimer's disease in rats, thereby significantly improving the learning and memory ability.     

胚胎神经干细胞移植治疗阿尔茨海默病

BACKGROUND:More recently, stem cell therapy has become an issue of concern. Exogenous neural stem cell transplantation brings new hope for the treatment of nervous system injury by self-replication and differentiation to complement and replace damaged or dead nerve cells. OBJECTIVE:To explore the therapeutic efficacy of neural stem cell transplantation on Alzheimer’s disease. METHODS:Thirty APP/PS1 mice with Alzheimer’s disease were randomly assigned into model group, cell solution transplantation group or cell transplantation group (n=10 per group). Another 10 C57BL/6 mice were selected as controls. Embryos of C57BL/6 mice at 18 embryonic days were taken to make neural stem cell suspension followed by transfection using lentiviral vectors carrying GFP gene at different multiplicities of infection (1, 5, 10, 15, 20). Afterwards, GFP-transfected neural stem cells were implanted into the hippocampus of Alzheimer’s disease mice in the cell transplantation group, while the same volume of complete medium was injected into the hippocampus of mice in the cell solution transplantation group. Morris water maze test was performed at 2 weeks after cell transplantation, and brain tissues of mice was taken and detected histologically at 4 weeks after cell transplantation. RESULTS AND CONCLUSION:Compared with the control group, the escape latency was significantly higher, and the number of crossings over the target quadrant was lower in the other three groups (P < 0.05). Compared with the cell solution transplantation and model groups, in contrast, the escape latency was significantly lower, and the number of crossings over the target quadrant was significantly higher in the cell transplantation group (P < 0.05). Four weeks after transplantation, more intact neurons were found in the cell transplantation group as compared with the model group. These findings indicate that neural stem cell transplantation can improve behavior and morphology performance of mice with Alzheimer’s disease.     

移植不同剂量脐带间充质干细胞提高老年痴呆大鼠学习记忆能力的比较

BACKGROUND:To delay the onset of Alzheimer’s disease, transplantation of viable and well-differentiated stem cells is expected to repair neural tissue, which has been an issue of concern. OBJECTIVE:To explore the effects of different doses of human umbilical cord mesenchymal stem cells (hUCMSCs) on learning and memory ability of Alzheimer’s disease rats. METHODS:Fifty Sprague-Dawley rats, 7 months of age, were randomized into normal, model, high-, middle- and low-dose hUCMSCs groups (n=10 per group). Rats in model and UCMSCs groups were used to make Alzheimer’s disease animals through intraperitoneal injection of 150 mg/kg D-galactose for 90 days, and rats in the normal group were given intraperitoneal injection of normal saline for 90 days. In the three hUCMSCs group, passage 3 hUCMSCs at doses of 1×10⁵/0.2 mL/20 g, 5×10⁵/0.2 mL/20 g, and 1×10⁶/0.2 mL/20 g were injected via the tail vein, respectively. Forty-five days after cell transplantation, Morris water maze test was used to detect rat’s learning and memory abilities, and hematoxylin-eosin staining was used to observe pathological changes of the rat hippocampal CA1 region. RESULTS AND CONCLUSION:Compared with the normal group, the rats in the model group showed significant reduction in the ability of learning and memory. Compared with the model group, the escape latency was significantly shortened in the middle-dose hUCMSCs group (P < 0.05), while the number of passing times through the platform was increased significantly (P < 0.05). In the model group, the cells in the hippocampal CA1 region were arranged irregularly with unclear nucleoli and a part of cells were concentrated and deeply stained. In the middle-dose hUCMSCs group, the cells in the hippocampal CA1 region were arranged regularly with clear nucleoli, and only individual cells were stained deeply. These findings indicate that middle-dose hUCMSCs transplantation can improve the learning and memory abilities of Alzheimer’s rats.     

骨髓间充质干细胞修复急性呼吸窘迫综合征

BACKGROUND:To date little is reported clinically on bone marrow mesenchymal stem cells (BMSCs) for acute respiratory distress syndrome (ARDS). OBJECTIVE:To study the effect of BMSCs in ARDS rats. METHODS:Sixty-three Sprague-Dawley rats were randomly assigned into normal control group, model and BMSCs groups (n=21 per group), followed by establishment of ARDS models. After awakening, rats in the model group were given no treatment, while those in the BMSCs group given tail vein injection of fluorouracil-labeled BMSCs. Then, pathological observation of the lung tissue was conducted using hematoxylin-eosin staining, and ELISA method was employed to detect interleukin-1β level in the rat lung tissue homogenates. Repair effects on lung injury were compared between two groups. RESULTS AND CONCLUSION:Hematoxylin-eosin staining showed that inflammation, edema and congestion in the rat lung tissue were vanished gradually, but no consolidation was found in the BMSCs group, while there was visible edema and congestion in the lung tissue of rats in the model group. Compared with the model group, pulmonary edema and interleukin-1β level in the lung tissue were significantly reduced in the BMSCs group (P < 0.05). Therefore, the whole bone marrow culture method is suitable to obtain BMSCs that have desired effects on ARDS in rats. In ARDS rats, due to inhalation of lipopolysaccharide, BMSCs can be directly involved in lung epithelial repair by reducing the release of inflammatory factors.     

骨髓间充质干细胞向软骨及骨分化：Wnt5a/PCP信号通路作用的研究与进展

BACKGROUND: Wnt5a is able to inhibit canonical Wnt signaling and activate non-canonical Wnt signaling pathway. In recent years, it has been found that non-classical Wnt5a/PCP signaling pathway mediated by Wnt5a plays an important role in the process of bone marrow mesenchymal stem cell proliferation and differentiation, but the underlying mechanism is unclear. OBJECTIVE: To summarize the progress in downstream effector molecules related to Wnt5a/PCP signaling pathway, and its roles in the chondrogenic and osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS: A computer-based online search of CqVip, CNKI and PubMed databases between January 2000 and February 2016 was performed using the Chinese keywords of “BMSCs, Wnt signaling pathways, chondrogenic differentiation, osteogenic differentiation” and English keywords of “BMSCs, chondrogenic differentiation, osteogenic differentiation, Wnt, Fzd, Ror2, RhoA, ROCK, JNK”, respectively. Literatures related to bone marrow mesenchymal stem cell chondrogenic and osteogenic differentiation were selected. Finally, 43 eligible articles were included for analysis through excluding the old and repeated research. RESULTS AND CONCLUSION:Wnt5a, a representative protein in non-canonical Wnt signaling pathway, paticipates in the cytoskeleton, cell migration and cell polarization and other activities by mediating its downstream signaling molecules such as Fzd, Ror, RhoA, ROCK, JNK, thereby regulating its proliferation and differentiation. But it is unclear how Wnt5a/PCP participates in the bone marrow mesenchymal stem cell chondrogenic and osteogenic differentiation and how the downstream effector molecules interact or function independently, which requires further studies. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

冠心病与非冠心病患者骨髓间充质干细胞基本生物学特性比较

BACKGROUND:It is unclear whether the body differences in patients with different types of heart diseases affect the characteristics and performance of stem cells. OBJECTIVE:To explore the biological characteristics of bone marrow mesenchymal stem cells from patients with different types of heart diseases. METHODS:Bone marrow mesenchymal stem cells were extracted using density gradient centrifugation from the bone marrow of 27 patients with coronary heart diseases and 20 patients with other heart diseases. Cell morphology was observed in the two groups. CD13, CD34, CD45, CD54, CD106 and CD44 positive expression was detected by flow cytometry. Cell proliferation was detected by MTT method, and the in vitro cell growth curves of the two groups were described. RESULTS AND CONCLUSION:The bone marrow mesenchymal stem cells of the two groups showed a long spindle shape, and there was no significant difference in the cell morphology between the two groups. In all the patients, the positive rates of CD34 and CD455 were less than 3.0%, while the positive rates of CD13 and CD44 were higher than 95.0%. However, the positive rates of CD54 and CD106 were higher in patients with coronary heart disease as compared with those with other heart diseases (P < 0.05). The in vitro growth curves of cells in the two groups were basically consistent, and the cell proliferation was only a little higher in the patients with other heart diseases compared with those with coronary heart disease. Experimental results show that different types of heart diseases in patients have no influence on morphology and proliferation of bone marrow mesenchymal stem cells, but some function-related proteins may exhibit certain difference in their expressions.     

骨髓间充质干细胞移植治疗老年性痴呆模型大鼠的行为学检测

BACKGROUND:More recently, studies have demonstrated that bone marrow mesenchymal stem cells can be induced in vitro to differentiate into neuron-like cells that are used for in vivo transplantation to repair nerve damage. OBJECTIVE:To study the effect of bone marrow mesenchymal stem cell transplantation on learning and memory ability of senile dementia rats. METHODS:Thirty male Sprague-Dawley rats were randomly divided into three groups: normal control group, stem cell therapy group and model control group. Rats in the latter two groups were used to establish animal models of senile dementia by intracranial injection of β-amyloid 1-40. Three weeks after modeling, rats were given bilateral hippocampal injection of induced bone marrow mesenchymal stem cell suspension in the stem cell therapy group, whereas no treatment was given in the normal control and model control groups. Morris water maze test was used to detect learning and memory ability of rats, and rat’s brain tissues were detected pathologically using hematoxylin-eosin staining. RESULTS AND CONCLUSION:After modeling, the escape latency was higher and the cross-platform frequency was lower in the model control group compared with the normal control group. After cell transplantation, the escape latency and cross-platform frequency were gradually shortened and increased with time, respectively. Compared with the model control group, the learning and memory abilities of rats were improved in the stem cell therapy group. The brain tissues were relatively intact in structure and exhibited less cell degeneration and necrosis in the stem cell therapy group compared with the model control group. To conclude, bone marrow mesenchymal stem cell transplantation exerts certain therapeutic effects on senile dementia by effectively improving the learning and memory ability.     

细胞传代对骨髓间充质干细胞向神经干细胞分化的影响

BACKGROUND:It is unclear whether serial cell passage in vitro influences the differentiation of bone marrow mesenchymal stem cells into neural stem cells. OBJECTIVE:To investigate the effect of cell passage on the differentiation of bone marrow mesenchymal stem cells into neural stem cells. METHODS:Rat bone marrow mesenchymal stem cells were isolated and cultured by the whole bone marrow adherence method. Bone marrow mesenchymal stem cells at passages 3, 6, 9, 12 were incubated in serum-free medium. After culture for 7 and 14 days, cell biological characterization was observed and differenitaiton ability into neural stem cells was observed by detecting Nestin expression in cells using flow cytometry. Then, the cells were further induced to differentiate and cell multipotential differentiation capacity was detected by measurement of nerve enolase and glial acidic protein expression. RESULTS AND CONCLUSION:Under induction, bone marrow mesenchymal stem cells at different passages were all differentiated into Nestin-positive neural stem cells. However, there was a significant difference in differentiation proportion of cells at different passages (P < 0.05). Strongest differentiation ability was found in the passage 6 cells, with the Nestin expression up to (93.7±2.3)% at 7 days of induction and (96.2±1.8)% at 14 days of induction. The proportion of differentiated cells at passages 6 and 9 was signfiicantly higher than that at passages 3 and 12. Moreover, adherent cells were positive for nerve enolase and glial acidic protein. All these findings indicate that the differentiation of bone marrow mesenchymal stem cells into neural stem cells is correlated with cell passage. Cells at lower or higher passages are both detrimental to cell differentiation.     

人参皂苷Rh2干预催产素诱导骨髓间充质干细胞向心肌细胞的转化

背景：课题组前期实验已证明了10 μmol/L催产素能诱导大鼠骨髓间充质干细胞向心肌细胞转化。 目的：观察人参皂苷Rh2在催产素诱导大鼠骨髓间充质干细胞向心肌细胞转化过程中的作用。 方法：采用贴壁法分离培养大鼠骨髓间充质干细胞。实验共分为5组,空白对照组细胞常规培养2周;催产素诱导组：10 μmol/L催产素连续诱导培养2周;人参皂苷Rh2低、中、高剂量组：分别加入0.5,1,2 μmol/L人参皂苷Rh2,培养24 h后加入10 μmol/L催产素,连续诱导培养2周。 结果与结论：光学显微镜下观察显示,与空白对照组相比,催产素诱导组的细胞部分细胞体积变大,部分细胞密集重叠生长,随人参皂苷Rh2剂量增大细胞密集重叠生长的范围增大。免疫组织化学染色和免疫印迹法结果显示,催产素诱导组和人参皂苷Rh2低、中、高剂量组中心肌肌钙蛋白T,连接蛋白43的蛋白表达均显著高于空白对照组(P < 0.05);人参皂苷Rh2剂量增大而阳性表达增强,并显著高于催产素诱导组(P < 0.05)。激光共聚焦检测结果显示,催产素诱导2周后,催产素诱导组骨髓间充质干细胞中游离钙的相对荧光强度显著升高(P < 0.05),而人参皂苷Rh2处理组的荧光强度高于催产素诱导组,与剂量呈正相关(P < 0.05)。结果证实,人参皂苷Rh2在体外可显著增强催产素诱导大鼠骨髓间充质干细胞向心肌细胞转化的作用。 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     

骨髓间充质干细胞联合茶黄素修复激素性股骨头坏死

背景：临床上对于股骨头坏死病理机制尚不完全知晓,大剂量使用糖皮质激素容易引起股骨头坏死。 目的：观察骨髓间充质干细胞联合茶黄素在大鼠激素性股骨头坏死中的临床治疗效果。 方法：体外分离培养大鼠骨髓间充质干细胞,将其复合在明胶海绵上。实验分4组,骨髓间充质干细胞联合茶黄素组采用液氮冷冻法建立激素性股骨头坏死鼠模型,并移植复合了骨髓间充质干细胞的明胶海绵,每天进行250 mg茶黄素灌服,设模型对照组,单纯减压组和细胞移植组作对照。 结果与结论：造模后4周,4组大鼠股骨头标本外形表现为圆形,关节软骨呈现为苍白色,关节软骨开始出现剥脱现象。造模后4周,模型对照组关节软骨剥脱加重,部分股骨头标本出现塌陷;骨髓间充质干细胞联合茶黄素组股骨头标本呈现出圆形,且为苍白色。造模后8周,模型对照组坏死区增加;单纯减压组出现成骨细胞,且存在纤维性骨痂形成;细胞移植组能够见到少量空骨陷窝,髓腔形态不规则;骨髓间充质干细胞联合茶黄素组出现大量新生骨形成,且髓内脂肪细胞规则。骨髓间充质干细胞联合茶黄素组造模后4,8周空骨陷窝阳性数显著少于其他3组(P < 0.05)。结果证实,激素性股骨头坏死采用骨髓间充质干细胞基础上联合茶黄素治疗效果理想。 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