骨髓间充质干细胞治疗吸入性肺损伤研究进展

成体干细胞来源广泛,无伦理争议,成为近几年关注热点.研究表明以骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)为代表的成体干细胞具有较强的多系分化潜能.骨髓间充质干细胞在肺损伤状态下可以向肺部移行,表现出肺上皮细胞的表型,参与损伤肺组织的修复重建,并可通过旁分泌途径等减轻肺损伤.并有研究表明干细胞的植入可以缓解和抑制损伤肺的病理进程.因此BMSCs移植为吸入性肺损伤的防治提供了一种崭新有效的方法.本文就BMSCs治疗热烟雾吸入性肺损伤中的应用综述如下.     

骨髓间充质干细胞在皮肤疾病治疗中的研究进展

骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BM-MSC)是一类来源于骨髓的非造血干细胞,它具有多向分化潜能和免疫调节作用,因为其获取容易,并能在体外扩增,使其成为应用最广泛的间充质干细胞。目前骨髓间充质干细胞在皮肤病的研究中受到广泛的关注,包括伤口愈合、放射性皮炎、系统性红斑狼疮(SLE)、硬皮病和银屑病等。     

银屑病患者骨髓间充质干细胞表面标志表达情况研究

目的研究银屑病患者骨髓间充质干细胞的体外分离培养及表面标志的表达情况,为银屑病患者间充质干细胞特性研究提供了基础。方法采用密度梯度离心法分离患者与对照组骨髓单一核细胞,通过贴壁法培养骨髓间充质干细胞,用流式细胞术鉴定细胞表面标志CD44、CD29、CD34、CD45、HLA-DR。结果经流式细胞仪测定的第三代培养的BMSCs表面标志CD44、CD29阳性表达,CD34、CD45、HLA-DR阴性表达。分析CD44、CD29,发现银屑病组明显低于对照组。结论密度梯度离心法结合贴壁法可获得较纯的BMSCs。银屑病患者表面标志CD44、CD29与正常人比较低表达。     

自体骨髓干细胞移植治疗糖尿病围手术期护理

近年来，随着对干细胞生物学特性认识和研究的不断深入，关于千细胞在治疗性血管新生中的作用越来越成为医学、生物学等方面关注的焦点。白体干细胞定向移植治疗糖尿病是近几年来周际上研究探索治疗糖尿病的一种新方法，通过重建胰岛功能，从根本上治疗糖尿病．有望控制或减少糖尿病一系列并发症的发生发展，提高生活质量。武警总医院自2009年9月应用自体骨髓干细胞移植治疗糖尿病，     

间充质干细胞与自身免疫性疾病

自身免疫性疾病的治疗一直以糖皮质激素和免疫抑制剂为主,但一些患者对糖皮质激素和免疫抑制剂反应不好或不能耐受,间充质干细胞可能成为一种新的辅助治疗手段.本文对间充质干细胞的特性及与自身免疫性疾病的关系进行综述.     

毛囊间充质干细胞的研究进展

 毛囊间充质干细胞（HFMSCs）是一类具有自我再生能力、高度增殖潜能、可多向分化且来源丰富的慢周期标记滞留细胞,可促进表皮、毛囊和皮脂腺再生。得益于其来源丰富、易获得、可于体外扩增、无需基因操作、不会形成肿瘤和无伦理限制等特点,毛囊间充质干细胞在再生医学中具有重要优势。Wnt、Shh、Notch和BMP等信号通路之间的协同和拮抗作用在干细胞稳态调节、表皮发育和毛囊再生过程中发挥至关重要的作用,这些途径的失调可能导致毛发脱落或者肿瘤的发生。本文综述毛囊间充质干细胞的分类及其特异性生物标记物、毛囊间充质干细胞的活化以及影响毛发再生的生物分子途径,为毛发疾病的治疗提供新的线索和靶点。     

体外诱导小鼠骨髓间充质干细胞分化为黑素细胞的观察

【摘要】 目的 探讨骨髓间充质干细胞体外诱导成为黑素细胞的可能性。方法　6周龄雄性C57BL/6小鼠股骨基质细胞行原代培养,6次传代后以氢化可的松、重组人胰岛素、转铁蛋白和成纤维细胞生长因子诱导黑素细胞。倒置光学显微镜观察细胞分化;透射电镜观察黑素小体成熟;免疫荧光染色观察黑素细胞相关表位表达;流式细胞仪检测黑素细胞的细胞周期及获得率。结果　6次传代间充质干细胞数量近109,免疫荧光检测CD44阳性率94.3%和CD105阳性率82.3%。培养180 d,细胞形态接近于黑素细胞,树突增多,胞质内出现黑素小体样结构,生长周期加快为3 ~ 4 d,肉眼可见棕黑色细胞沉淀。电镜观察显示Ⅳ期为主的黑素小体。免疫荧光显示酪氨酸酶相关蛋白-1,酪氨酸酶相关蛋白-2和小眼畸形相关转录因子阳性。流式细胞仪分析显示细胞基本处于G1和S期。酪氨酸酶相关蛋白-1阳性的黑素细胞获得率约为80%。结论　骨髓间充质干细胞可以被大量诱导分化为黑素细胞;诱导黑素细胞的形态学、超微结构、特异性表位等皆接近于正常黑素细胞,具有一定的增殖活性,获得率较高。 【关键词】 间质干细胞; 骨髓; 黑素细胞; 体外研究     

HaCaT细胞诱导兔骨髓间充质干细胞表达角蛋白的初步研究

目的:探讨在体外培养的条件下HaCaT细胞诱导兔骨髓间充质干细胞(MSCs)表达角蛋白的可能性。方法:体外分离培养MSCs,流式细胞术检测CD34、CD45和CD44蛋白的表达以鉴定MSCs细胞。传代后与HaCaT共培养。分别在共培养3、7天后,密度梯度离心法分离MSCs和HaCaT,免疫细胞化学方法检测角蛋白Pan-cytokeratin(Pan-CK)、CK19的表达。结果:流式细胞术检测体外培养的MSCs CD34、CD45阴性,CD44阳性,表明MSCs培养成功。MSCs与HaCaT共培养3天、7天后,免疫细胞化学检测结果显示Pan-CK和CK19均为阳性。共培养7天的MSCs Pan-CK阳性率明显高于共培养3天的MSCs[分别为(29.1±8.3)%、(13.5±3.7)%,t=13.35,P0.05],共培养7天的MSCs CK19阳性率与共培养3天的MSCs相比无统计学差异[分别为(3.3±0.8)%、(3.1±0.5)%,t=1.37,P0.05]。结论:与HaCaT细胞共培养可诱导兔骨髓间充质干细胞向表皮细胞及表皮干细胞分化。     

HaCaT细胞诱导兔骨髓间充质干细胞表达角蛋白的初步研究

目的:探讨在体外培养的条件下HaCaT细胞诱导兔骨髓间充质干细胞(MSCs)表达角蛋白的可能性。方法:体外分离培养MSCs,流式细胞术检测CD34、CD45和CD44蛋白的表达以鉴定MSCs细胞。传代后与HaCaT共培养。分别在共培养3、7天后,密度梯度离心法分离MSCs和HaCaT,免疫细胞化学方法检测角蛋白Pan-cytokeratin(Pan-CK)、CK19的表达。结果:流式细胞术检测体外培养的MSCs CD34、CD45阴性,CD44阳性,表明MSCs培养成功。MSCs与HaCaT共培养3天、7天后,免疫细胞化学检测结果显示Pan-CK和CK19均为阳性。共培养7天的MSCs Pan-CK阳性率明显高于共培养3天的MSCs[分别为(29.1±8.3)%、(13.5±3.7)%,t=13.35,P<0.05],共培养7天的MSCs CK19阳性率与共培养3天的MSCs相比无统计学差异[分别为(3.3±0.8)%、(3.1±0.5)%,t=1.37,P>0.05]。结论:与HaCaT细胞共培养可诱导兔骨髓间充质干细胞向表皮细胞及表皮干细胞分化。     

兔骨髓间充质干细胞低氧培养及基本性质研究

目的探讨体外低氧浓度培养条件对于新西兰大白兔骨髓问充质干细胞（bone marrow mesenchymal stem cells，BM—MSCs）增殖、分泌等特性的影响。方法将细胞分为低氧浓度培养组及对照组，分别检测其细胞表面标志物表达、分化能力、生长曲线、克隆形成能力，并使用ELISA法检测两组胶质细胞源性神经营养因子（GDNF）分泌量的区别。结果低氧浓度培养组细胞在表面标志物表达及分化能力上均与对照组无明显区别，而增殖能力及克隆形成能力较对照组明显增强，经ELISA检测，低氧浓度培养条件下的细胞GDNF的分泌量更高。结论低氧浓度的培养条件有利于BM—MSCs增殖及分泌能力增强，适宜用于BM—MSCs的扩增。     

皮肤间充质干细胞的原代培养

目的评估体外分离培养皮肤间充质干细胞(sMSCs)的适宜方法。方法采用低血清培养和无血清培养两种不同的原代培养法,体外培养皮肤间充质干细胞,通过观察细胞形态特征及流式细胞技术进行鉴定,比较其差异。结果两种培养法均能培养出形态均一、成熟的皮肤间充质干细胞,但是低血清法培养细胞所用时间8.93d明显小于无血清培养法所需时间15.93d,差异有统计学意义(P<0.001),且低血清法培养出的细胞较无血清培养方法得到的皮肤间充质干细胞的CD44阳性率较高,CD45阴性率较低,差异有统计学意义(P<0.001)。结论低血清培养法是一种适用于临床试验研究皮肤间充质干细胞的原代培养方法。     

间充质干细胞与黑素细胞共培养的研究进展

自体黑素细胞移植为稳定期白癜风的一线治疗手段。该治疗虽已被证实复色有效,但是如何维持移植细胞的未分化状态、提高移植细胞的活力、增加其在移植区的附着度,最大程度实现复色目的尚需进一步研究。既往研究多局限于单纯黑素细胞或黑素细胞与角质形成细胞共培养,但获得足够数目的自体角质形成细胞会遗留大面积瘢痕。间充质干细胞是目前临床研究的理想种子细胞,来源丰富、获取方便,与黑素细胞共培养有望促进移植区复色。本文对间充质干细胞与黑素细胞共培养的研究进展做一综述。     

小鼠真皮间充质干细胞与成纤维细胞共培养对胶原分泌和TGF-β1表达的影响

目的 探讨真皮间充质干细胞在皮肤组织修复中的作用.方法 采用低血清培养基,消化-贴壁-传代法体外培养、鉴定小鼠真皮间充质干细胞(mdMSC),并与体外分离培养的正常人皮肤成纤维细胞于transwell小室培养体系中共培养,样本碱水解法和ELISA法分别检测第4、8天培养上清液中羟脯氨酸和TGF-β1的变化.结果 共培养第8天,经mdMSC 2.5×104和mdMSC 1×104处理的正常人皮肤成纤维细胞培养上清液中羟脯氨酸含量较单独培养时明显增高(P<0.05).经mdMSC处理的各组正常人皮肤成纤维细胞培养上清液中TGF-β1含量于共培养第8天时均高于单独培养(P<0.01);经mdMSC 1×104处理的正常人皮肤成纤维细胞培养上清液中TGF-β1含量在第4天亦高于单独培养,差异有统计学意义(P<0.05).各不同细胞密度的MSC处理组的羟脯氨酸含量与TGF-β1水平无相关关系(r=0.108,P＞0.05).结论 mdMSC与正常人皮肤成纤维细胞共培养可增加羟脯氨酸和TGF-β1的分泌,可能是mdMSC促进皮肤组织修复的机制之一.     

小鼠真皮间充质干细胞与成纤维细胞共培养对胶原分泌和TGF-β1表达的影响

Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.     

小鼠真皮间充质干细胞与成纤维细胞共培养对胶原分泌和TGF-β1表达的影响

Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.     

小鼠真皮间充质干细胞与成纤维细胞共培养对胶原分泌和TGF-β1表达的影响

Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.     

小鼠真皮间充质干细胞与成纤维细胞共培养对胶原分泌和TGF-β1表达的影响

Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.     

小鼠真皮间充质干细胞与成纤维细胞共培养对胶原分泌和TGF-β1表达的影响

Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.     

小鼠真皮间充质干细胞与成纤维细胞共培养对胶原分泌和TGF-β1表达的影响

Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.     

小鼠真皮间充质干细胞与成纤维细胞共培养对胶原分泌和TGF-β1表达的影响

Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.