首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 62 毫秒
1.
人骨髓间质干细胞分离和培养扩增方法的研究   总被引:3,自引:0,他引:3  
目的 :探索分离、纯化及培养成人骨髓间质干细胞 (hMSC)的最佳方法。方法 :采用Ficoll Paque淋巴细胞分离液和全血接种两种方法分离成人MSC ,筛选体外培养hMSC适合的培养基和适宜的血清含量 ,流式细胞仪检测hMSC表面抗原表达。结果 :经Ficoll Paque淋巴细胞分离液分离后 ,接种于 φ =10 0mL/L胎牛血清的L DMEM培养液 ,细胞贴壁良好 ,增殖速度快 ,hMSC在体外扩增 5代可获得 1× 10 8细胞。全血接种分离hMSC ,细胞贴壁慢、少 ,生长情况欠佳。除L DMEM ,其他类型的培养基均不适合hMSC的培养扩增。流式细胞仪检测结果显示CD2 9、CD44、CD10 5、CD166表达阳性 ,CD14、CD3 3、CD3 4、CD3 8、CD45、CD11a为阴性。结论 :用Ficoll Paque淋巴细胞分离液分离骨髓单核细胞 ,接种于 φ =10 0mL/LFCS的L DMEM培养液 ,分离培养扩增hMSC的效果最好  相似文献   

2.
丹参注射液诱导间质干细胞分化为神经元样细胞   总被引:4,自引:0,他引:4  
[目的]丹参注射液体外定向诱导成人骨髓间质干细胞(MSC)分化为神经元样细胞.[方法]采用Ficoll-Paque液离心分离成人MSC,体外扩增,流式细胞仪检测MSC表面抗原表达,分别采用含丹参注射液或硫代甘油等试剂的无血清达乐伯克改良必需基本培养基(DMEM)诱导MSC分化为神经元,免疫组化鉴定神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF-M)、胶质纤维酸性蛋白(GFAP)、巢蛋白(nestin)的表达.[结果]成人骨髓间质干细胞在体外扩增原代可获得0.5×106,15代可获得(2~3)×1012个细胞,细胞流式细胞仪检测结果显示CD29、CD44、CD90、CD105、CD166表达阳性,CD11a、CD14、CD34、CD38、CD45、CD80、CD86为阴性.加入丹参或硫代甘油诱导后,MSC胞体收缩,突起伸出;免疫组化显示诱导出的神经元样细胞NSE、NF-M、nestin表达阳性,GFAP阴性.[结论]丹参可以在体外诱导成人骨髓间质干细胞分化为神经元样细胞.  相似文献   

3.
成人骨髓间充质干细胞的体外培养和表型鉴定   总被引:6,自引:0,他引:6  
目的 建立成人骨髓间充质干细胞(hMSCs)的分离培养方法,并对其表面标志进行检测,研究骨髓间质干细胞(MSC)基本生物学特性。方法 采用Percoll(1.073 g/mL)梯度离心分离hMSCs,体外扩增,流式细胞仪检测hMSCs表面抗原表达。结果成人骨髓间质干细胞在体外扩增原代及传代培养显示,hMSCs具有活跃增殖的能力,每10 mL骨髓可获得(1.25±0.56)×106个原代细胞,7代可获得(5.32±1.15)×1010个细胞(n=5),随着传代次数的增加,细胞的增殖能力下降。流式细胞仪检测生长良好的第3代hMSC显示CD29、CD44、CD106、CD105、CD166、HLA-A/B/C表达阳性,CD34、CD45、CD14、CD31、CD49d、CD54、HLA-DR/DP/DQ表达为阴性。细胞周期分析显示,hMSCs中,S+G2+M期的细胞约占(16.25±3.18)%,其中s期为(4.12±2.35)%;而处于G0+G1期的细胞总数占(76.35±5.28)%,说明大部分细胞仍然处于静止期。流式细胞仪检测表明分离的hMSCs具有间充质干细胞的特征。结论 hMSCs有很强的自我更新能力,在体外传代培养可维持良好的干细胞生物学特性,为进一步深入研究hMSCs作为组织工程种子细胞的应用提供参数。  相似文献   

4.
目的建立成人骨髓间充质干细胞(hMSCs)的分离培养方法,并对其表面标志进行检测,研究骨髓间质干细胞(MSC)基本生物学特性.方法采用Percoll(1.073 g/mL)梯度离心分离hMSCs,体外扩增,流式细胞仪检测hMSCs表面抗原表达.结果成人骨髓间质干细胞在体外扩增原代及传代培养显示,hMSCs具有活跃增殖的能力,每10 mL骨髓可获得(1.25±0.56)×106个原代细胞,7代可获得(5.32±1.15)×1010个细胞(n=5),随着传代次数的增加,细胞的增殖能力下降.流式细胞仪检测生长良好的第3代hMSC显示CD29、CD44、CD106、CD105、CD166、HLA-A/B/C表达阳性,CD34、CD45、CD14、CD31、CD49d、CD54、HLA-DR/DP/DQ表达为阴性.细胞周期分析显示,hMSCs中,S+G2+M期的细胞约占(16.25±3.18)%,其中S期为(4.12±2.35)%;而处于G0+G1期的细胞总数占(76.35±5.28)%,说明大部分细胞仍然处于静止期.流式细胞仪检测表明分离的hMSCs具有间充质干细胞的特征.结论 hMSCs有很强的自我更新能力,在体外传代培养可维持良好的干细胞生物学特性,为进一步深入研究hMSCs作为组织工程种子细胞的应用提供参数.  相似文献   

5.
【目的】 从成人骨髓中分离出Muse细胞,体外诱导成神经前体细胞,为细胞移植修复神经系统损伤提供种子细胞。 【方法】 利用密度梯度离心和差速贴壁法从健康成人骨髓中分离、培养骨髓基质干细胞,通过免疫荧光细胞化学技术和流式细胞仪技术对其进行鉴定;体外扩增、传代6次后利用Muse细胞特征性的SSEA-3/CD105分子表型阳性,通过流式细胞仪从骨髓基质干细胞中分选出Muse细胞并进行体外培养,观察其干细胞成球特性;对Muse细胞进行胰蛋白酶孵育,分析其应激耐受能力;利用免疫荧光细胞化学技术和RT-PCR技术检测Muse细胞的多能干细胞标记物表达情况;通过在培养基中添加成纤维细胞生长因子、表皮生长因子等对Muse细胞向神经前体细胞方向进行体外诱导,观察神经前体细胞的干细胞成球情况,利用免疫荧光细胞化学技术和RT-PCR技术对其进行表型鉴定并观察神经前体细胞标记物的表达情况。 【结果】 从成人骨髓中分离出骨髓基质干细胞,免疫荧光细胞化学检测和流式细胞仪检测结果显示CD90表达阳性、CD45和CD11b表达阴性;通过流式细胞仪从骨髓基质干细胞中分选出约0.5%的Muse细胞进行培养,细胞聚集成球,悬浮生长,表现为胰蛋白酶耐受;免疫荧光细胞化学检测和RT-PCR检测结果显示Muse细胞表达多能干细胞标记物Nanog、Oct4、Sox2阳性;体外诱导Muse细胞向神经前体细胞方向分化,观察到细胞成球现象,免疫荧光细胞化学检测和RT-PCR检测结果显示诱导后细胞表达神经前体细胞标记物nestin、βIII-tubulin。 【结论】 从成人骨髓中成功分离出Muse细胞,具有多能干细胞特性,经体外诱导形成神经前体细胞,为以后组织工程移植修复神经损伤提供新的种子细胞。  相似文献   

6.
定向诱导兔骨髓间质干细胞分化为神经元样细胞的研究   总被引:2,自引:0,他引:2  
目的 :体外定向诱导兔骨髓间质干细胞 (MSC)分化为神经元样细胞。方法 :采用Ficoll Paque液 (1 .0 83g/L)离心分离兔MSC ,体外扩增 ,硫代甘油等试剂的无血清达乐伯克改良必需基本培养基 (DMEM)诱导MSC分化为神经元。免疫组化鉴定神经元烯醇化酶 (NSE)、神经丝蛋白 (NF)、胶质纤维酸性蛋白 (GFAP)的表达。结果 :兔骨髓间质干细胞在体外扩增原代可获得 5× 1 0 5、1 0代可获得 2× 1 0 1 0 个细胞。加入硫代甘油诱导后 ,MSC胞体收缩 ,突起伸出 ;免疫组化显示诱导出的神经元样细胞NSE、NF、表达阳性 ,GFAP阴性。结论 :兔骨髓间质干细胞在体外可以分化为神经元样细胞  相似文献   

7.
人脐带间质干细胞分离纯化及基本生物学特性研究   总被引:4,自引:0,他引:4  
目的探讨人脐带间质干细胞(MSC)的分离、纯化、扩增方法,研究其基本生物学特性.方法无菌条件下收集剖宫产新生儿脐带,分别用组织块贴壁法和酶消化法获取脐带间质干细胞,进行原代培养.应用DMEM/F12培养液进行纯化和扩增培养.用流式细胞仪检测MSC的细胞表面标志.绘制细胞生长曲线并测定细胞周期.结果两种分离方法均可获得MSC,原代培养12~14 d后可达90%融合,细胞可传代20代以上.流式细胞仪检测结果显示,脐带MSC强表达CD13、CD29、CD44、CD105,弱表达CD106,不表达CD34、CD11a、CD14、CD31、CD45.结论脐带MSC在体外有较强的增殖能力,可作为组织工程的种子细胞.  相似文献   

8.
成人骨髓间充质干细胞体外培养扩增及其生物学特性研究   总被引:6,自引:0,他引:6  
罗依  梁彬  余勤 《浙江医学》2003,25(8):468-469,473
目的 建立成人骨髓间充质干细胞 (MSCs)体外培养和扩增的条件 ,探讨其生物学特性。方法 取正常成人骨髓用含10%胎牛血清的LG -DMEM培养液培养、扩增后 ,进行透射电镜观察 ,流式细胞仪行细胞表面抗原及细胞周期分析检测。结果 培养扩增获取的成人骨髓MSCs形态均一 ,增殖能力强 ,细胞扩增至第5代细胞数达 (2~3)×108。该类细胞CD29、CD44、CD166表达阳性 ,CD14、CD34、CD45、HLA -DR表达阴性。细胞周期以G0/G1 期为主。细胞胞浆具丰富粗面内质网。结论 所建立的分离和培养方法可获取骨髓粘附细胞中一组独特的细胞群 ,具有MSCs的生物学特性。  相似文献   

9.
不同来源的人间质干细胞分离与基本生物学特性研究   总被引:3,自引:2,他引:1  
目的:分离并比较人胚胎骨髓、成人骨髓和新生儿脐血来源间质干细胞(MSC)的生物学特性,为组织工程选取种子细胞提供实验依据.方法: 用贴壁方法分离三种来源的间质干细胞,观察其形态与生长曲线,并用流式细胞仪测定其表面标志.结果: 骨髓标本均可成功分离出MSC,但是人胚胎骨髓MSC增殖能力要明显强于成人骨髓MSC,而脐血分离MSC没有得到成功.结论: 人胚胎骨髓MSC是很好的组织工程种子细胞,成人骨髓MSC的增殖能力相对较弱,应用受到一定的限制.脐血MSC分离与扩增有待进一步研究.  相似文献   

10.
【目的】研究体外培养骨髓来源的表达nestin的成年大鼠骨髓间质干细胞(MSC)并诱导分化神经元样细胞的潜能,为中枢神经系统疾病的细胞移植治疗提供理想的供体。【方法】采用全骨髓法分离培养成年SD大鼠MSC,传至第6代的MSC接种在前一天涂有100ug/L多聚赖氨酸的塑料培养瓶或培养皿中,用含100mL/LFBS的DMEM/F12培养液添加10ng/mLbFGF的培养液培养并以DMSO、BHA与Forskolin等诱导剂联合诱导MSC分化为神经元。流式细胞仪鉴定诱导前MSC的细胞表面抗原,免疫荧光检测诱导前及诱导后6h、12h、24h神经元特异性抗原nestin、NF-200和GFAP表达率。【结果】流式细胞仪检测传至第6代MSC表面抗原CD29和CD44阳性率分为98.8%,96.6%,CD34和CD45阴性。免疫荧光鉴定传至第6代的MSC表达nestin阳性率为57.1%±6.9%,不表达NF-200和GFAP。诱导后30min可见神经元样细胞出现,诱导后6h、12h、24h经nestin和NF.200免疫荧光鉴定,诱导后6h、12h、24hnestin阳性率分别为96.5%±1.9%,88.1%±5.4%,33.5%±5.4%,NF-200阳性率分别为90.1%±2.9%,97.5%±1.3%,98.1%±1.6%。【结论】骨髓来源的nestin阳性的MSC具有神经祖细胞特性并且可诱导分化为神经元样细胞,可作为种子细胞用于神经系统疾病的治疗。  相似文献   

11.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

12.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

13.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

14.
Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.  相似文献   

15.
Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.  相似文献   

16.
A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care(~'4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.  相似文献   

17.
Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.  相似文献   

18.
目的:评价使用安心颗粒对急诊经皮冠状动脉介入术(PPCI)术后生活质量的影响.方法:将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组(术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次)和对照组(仅接受基础药物治疗).所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表(MIDAS)、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果:入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异(P>0.05);出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低(P<0.05);组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低(P<0.05).两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血(P>0.05).两组发生轻度血小板减少症的患者数比较无差异(P>0.05).结论:PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.  相似文献   

19.
Journal of Nanjing Medical University (English Edition) JNMU, sponsored by Nanjing Medical University, was established in 1987. It is a bimonthly comprehensive English medical journal published locally and abroad.Since 2007, Journal of Nanjing Medical University (English Edition )was granted Elsevier the full publishing and distribution rights worldwide for the Electronic Edition, excluding the People's Republic of China.  相似文献   

20.
Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P<0.01);the MAP decreased(P<0.01)and the HR increased drastically(P<0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号