骨髓间充质干细胞分离培养和定向分化的研究现状

摘要：骨髓间充质干细胞是存在于骨髓基质中的非造血系细胞，它们能够分化成中胚层和非中胚层细胞。骨髓间充质干细胞主要有3种分离方法：贴壁法，梯度离心法和免疫选择法。文章列举了在特定的诱导条件下骨髓间充质干细胞的定向分化种类，讨论了文献报道的各种相关的诱导条件，分析这些人工调控条件后的原理并探寻统一而有效的诱导条件。得出：骨髓间充质干细胞在适当条件下不仅可以分化为同源于中胚层的间质组织细胞，还可以突破胚层界限，分化为非中胚层组织，如脂肪细胞、骨细胞、软骨细胞等。作为组织工程的种子细胞及细胞替代治疗的一种来源，骨髓间充质干细胞被广泛应用于干细胞的研究中。     

脂肪间充质干细胞在组织工程中的应用

背景：脂肪间充质干细胞与骨髓间充质干细胞具有相似的形态学及生物学特性,可作为组织工程的种子细胞来源。 目的：深入认识脂肪间充质干细胞的生物学特性,对其在组织工程中的研究进展及临床应用、基础研究前景进行探讨。 方法：以adipose tissue-derived stem cell,tissue engineering,stem cells为检索词,检索Pubmed 数据库(1999-01/ 2009-06)。以脂肪间充质干细胞,组织工程为检索词,检索CNKI期刊全文数据库(1999-01/2008-12)。文献检索语种限制为英文和中文。以脂肪间充质干细胞成骨和软骨的能力以及脂肪间充质干细胞和基因转染技术联合治疗疾病的效果为评价指标。纳入大鼠脂肪和骨髓来源间充质干细胞成骨分化比较的体外研究,排除重复性研究及目的与本文不相关的研究。 结果与结论：脂肪间充质干细胞取材方便,来源丰富,可在体外稳定增殖传代,与骨髓间充质干细胞具有相似的形态学及生物学特性,在一定诱导条件作用下可以定向分化为中胚层及内、外胚层组织细胞。脂肪间充质干细胞联合组织工程学支架在一定程度上可以修复骨、关节软骨的缺损,但新生软骨的质量与周围软骨的连接、生物力学强度、未来退化等都与正常的软骨有一定差距,尚需深入研究。随着人们对成体间充质干细胞研究的不断深入,发现因间充质干细胞具有自我扩增和分化潜能,使导入的“外源基因”能够有效扩散;获取和体外扩增比较容易,可在体外对其进行基因改造和修饰,且其易被外源基因转染。因此,脂肪来源的间充质干细胞可以结合基因工程学,应用于基因治疗。但是在当前研究方面,尚存在基因载体潜在致癌性,转染后对人体及干细胞是否产生不良影响等问题尚未阐明。     

人胎盘源与人骨髓源间充质干细胞的生物学性状比较

人骨髓源间充质干细胞具有很好的临床应用价值,但数量和取材都有限,而人胎盘源间充质干细胞则相反,目前已成为再生医学的重要细胞来源和最具临床应用前景的功能干细胞。 目的：比较人胎盘源间充质干细胞和人骨髓源间充质干细胞体外分离培养、扩增及生物学性状的差异。 方法：采用胶原酶消化法分离人胎盘组织,密度梯度离心法分离骨髓单个核细胞,分别加入体积分数10%胎牛血清的LG-DMEM 培养液,待细胞汇合至90%后消化传代。分别取第3代的人胎盘和骨髓间充质干细胞,按1×106 浓度接种,当细胞达70%~80%融合时,换成成脂细胞诱导分化培养液,诱导16 d。 结果与结论：胎盘间充质干细胞、骨髓间充质干细胞均成贴壁生长、形态均一的成纤维样细胞梭形外观,但后者体积略小。两种细胞均高表达CD29、CD44,不表达CD34、CD106。二者均可分化为脂肪细胞。可见,从胎盘和骨髓中培养出的间充质干细胞在细胞形态、生长特性等方面基本相似,在体外均可有效扩增并成脂肪分化,均可作为组织工程的另一成体干细胞来源,而胎盘源间充质干细胞具有更好的应用前景。     

间充质干细胞在组织工程中的应用进展***☆

背景：间充质干细胞主要存在于结缔组织和器官间质中,具有强大的增殖能力和多向分化潜能及免疫调节等多种功能;同时具有来源方便,易于分离、培养、扩增和纯化等优点,因而在组织工程,基因治疗等领域日益受到重视。 目的：综述不同来源间充质干细胞的生物学特性、对其他细胞的支持作用、免疫原性、免疫调节作用及其应用。 方法：应用计算机检索2001-03/2010-03 PubMed数据库相关文章,检索词为“mesenchymal stem cells,tissue engineering”,同时检索万方数据库相关文章,检索词为“间充质干细胞,组织工程”。共检索到文献182篇。 结果与结论：间充质干细胞来源于中胚层间充质,不仅具有多向分化的潜能,单独应用于组织工程,而且具有为其他细胞的生长繁殖提供支持的作用,可以调节机体的免疫状态等特性而被广泛应用于医学领域。目前,间充质干细胞功能的研究虽然取得了一些成就,但有关研究才刚刚起步,仍存在许多理论和技术问题,对于如何有效控制其在体内定向分化、增殖、迁移并整合参与宿主细胞组织功能还需要大量深入和系统研究。     

成体干细胞向血管内皮细胞定向分化的研究现状

成体干细胞存在于已分化的组织器官中，具有明显的可塑性，能跨系统、跨胚层分化，在组织工程和损伤修复领域具有重要的应用价值。在血管组织工程研究领域中，成体干细胞作为种子细胞得到越来越广泛而深入的研究。目前研究表明，具有向血管内皮细胞分化的成体干细胞主要有骨髓间充质干细胞、造血干细胞、脂肪来源的干细胞以及羊膜来源的干细胞等。基于此，就成体干细胞作为种子细胞在血管组织工程领域研究现状进行综述。     

大鼠骨髓间充质干细胞及大隐静脉来源细胞向组织工程化韧带种子细胞诱导的比较

背景：关节病中韧带损伤难以治愈，且传统的韧带替代物有很多不足。利用组织工程方法治疗关节病，需要选择适合的种子细胞。 目的：观察诱导分化SD大鼠骨髓间充质干细胞及大隐静脉来源细胞作为韧带组织工程种子细胞的可行性。 方法：胰酶-去垢剂法制备去细胞韧带组织；密度梯度离心和贴壁细胞培养相结合的方法体外分离扩增大鼠骨髓间充质干细胞，成纤维细胞生长因子诱导其分化为成纤维细胞，设大隐静脉间质细胞为对照，比较2种细胞形态学、增殖能力、胶原合成以及在韧带上的生长情况方面的异同。 结果与结论: 大鼠骨髓间充质干细胞及大隐静脉来源细胞均为贴壁生长，形态为梭形或多角型，两者的冻存和复苏率、合成胶原能力方面无显著差异，都能种植在韧带组织上并在其上面生长。骨髓间充质干细胞诱导分化的成纤维细胞与静脉来源的间质细胞之间无显著差异。 关键词：大隐静脉；骨髓间充质干细胞；种子细胞；组织工程；诱导分化；韧带；大鼠     

骨髓间充质干细胞治疗缺血性脑卒中的机制

骨髓间充质干细胞(bone marrow mesenchymal stemcells,BMSCs)是间充质来源的多潜能干细胞,可以促进损伤、衰老器官的结构及功能修复,是当前再生医学最有潜力的种子细胞.BMSCs在体外具有分化为肌肉、骨骼、脂肪及神经等多种组织前体细胞的潜能,近年来国内外对BMSCs治疗缺血性脑卒中的研究较多[1-3].     

胎盘来源间充质干细胞的生物学特征

背景：近几年间充质干细胞的研究取得了一定的进展,但仍存在来源匮乏及免疫排斥等问题。胎盘间充质干细胞具有诸多优点可能成为间充质干细胞新的来源。 目的：分析总结胎盘间充质干细胞分离、培养及诱导分化的研究进展。 方法：由该论文的研究人员应用计算机检索PubMed数据库1993/2009有关胎盘间充质干细胞性质及诱导分化的文献,检索词“mesenchymal stem cells,placenta”,共检索到180 篇文献,对资料进行初审,排除重复性研究。所选用的41篇文章中,3篇为综述,其余均为临床或基础实验研究。 结果与结论：与研究比较多的骨髓间充质干细胞相比,胎盘间充质干细胞因具有免疫原性低、取材方便、无侵入性操作、无伦理道德限制、分化潜能大、增殖能力强、易于工业化制备等特点,成为再生医学的重要细胞来源和最具临床应用前景的功能干细胞。     

间充质干细胞的生物学特性及多向分化潜能*★

学术背景：间充质干细胞是造血微环境中的一种重要细胞成分，可以向多种组织增殖分化，且免疫原性低，临床实验证实间充质干细胞移植可用于组织修复及治疗间充质组织遗传缺陷性疾病。 目的：深入认识间充质干细胞的生物学特性及其多向分化潜能。 检索策略：由该论文的研究人员应用计算机检索Pubmed数据库1995-01/2005-12的相关文献，检索词“mesenchymal stem cells，differentiation，biological character”，并限定文章语言种类为English。同时计算机检索中国期刊全文数据库1995-01/2005-12的相关文献，检索词“间充质干细胞，多向分化，生物学性质”，并限定文章语言种类为中文。共检索到78篇文献，对资料进行初审，纳入标准：①文章所述内容应与间充质干细胞的生物学特性或多向分化潜能密切相关。②同一领域选择近期发表或在权威杂志上发表的文章。排除标准：①重复性研究。②Meta分析。 文献评价：文献的来源主要是通过对间充质干细胞的生物学特性及其多向分化潜能方面内容进行汇总分析。所选用的30篇文献中，4篇为综述，其余均为临床或基础实验研究。 资料综合：①间充质干细胞在骨髓组织中的含量最为丰富，此外还存在于胎盘、羊水、脐静脉内皮下层、外周血及肝脏、脂肪、肌肉、皮肤等多种组织中。间充质干细胞具有高度增殖、自我更新和多向分化潜能，在不同诱导条件下可分化为软骨、骨、骨骼肌、肌腱、脂肪、神经及肾脏实质的细胞等。②间充质干细胞体外培养的方法目前主要有3种：密度梯度离心与贴壁筛选法：鉴于间充质干细胞与其他细胞密度的差异，采用Percoll或者Ficoll分离液来实现细胞间的分离，同时依据间充质干细胞贴壁生长的特性，将其与非贴壁细胞分离。流式细胞仪分选法：根据间充质干细胞体积小、相对缺少颗粒这一特性采用流式细胞仪对其进行分选。磁珠分选法：根据间充质干细胞表面带有或缺失的抗原成分进行正选或负选，用抗体包被磁珠，获得相对纯化的间充质干细胞。现今常用含5%~20%胎牛血清的DMEM培养基培养间充质干细胞。目前对于间充质干细胞的鉴定也主要是根据形态、功能等，而且不同来源的间充质干细胞表面标志物也不尽相同。③间充质干细胞在特定的理化环境和细胞因子诱导下，能够分化为成骨细胞、脂肪细胞、神经细胞、心肌细胞、内皮等多种细胞类型，受到很多因素的影响，而且免疫原性弱，是组织工程理想的种子细胞来源。此外，间充质干细胞易于外源基因的转染和表达，在细胞治疗和基因治疗中也有着广泛的应用。 结论：间充质干细胞的多向分化特性使其在组织工程创伤修复、细胞替代治疗、支持造血、基因治疗等方面的应用前景相当广阔。但目前采用的体外分离培养无法得到单克隆的细胞成分，也缺乏可靠的鉴定标准，不同来源的间充质干细胞有何差异等问题亟待解决。     

骨髓间充质干细胞在软骨缺损修复中的应用

学术背景：骨髓间充质干细胞由于其易于获取,并可以在体外短期内大量扩增,是目前最有希望应用于软骨组织工程的干细胞。 目的：对骨髓间充质干细胞在软骨缺损修复中的应用状况进行综述。 检索策略： 由该论文的研究人员应用计算机检索1982-10/2006-12 Pubmed数据库与骨髓间充质干细胞修复软骨缺损相关文献，检索词“Marrow; Mesenchymal stem cells；cartilage defect”，限定语言种类为English, 同时检索1982-10/2006-12 中国科技成果数据库检索词“骨髓；间充质干细胞；软骨缺损”，并限定文章语言种类为中文。共检索到126篇文献，对资料进行初审，纳入标准：①综述类及实验类文章；②中文核心期刊收录文献。排除标准：重复性研究。 文献评价：文献的来源主要来自Pubmed数据库及中国科技成果数据库。文献类型为综述类及实验研究。 资料综合：骨髓间充质干细胞在体内具有分化产生软骨的能力已被证明,而在体外培养条件下软骨表型的分化却是一个受多种因素限制的复杂过程,目前调控机制仍不明确。动物实验表明,骨髓间充质干细胞能够修复具有临床意义的骨和软骨缺损。虽然近年来对骨髓间充质干细胞及其在软骨组织工程方面的研究已取得较大进展,但对骨髓间充质干细胞分化各阶段细胞标志物、骨髓间充质干细胞的增殖、分化的控制及基因转染技术及临床应用的评估结果都有待进一步研究。 结论：动物实验表明,骨髓间充质干细胞能够修复具有临床意义的骨和软骨缺损，虽然近年来对骨髓间充质干细胞及其在软骨组织工程方面的研究已取得较大进展,但对骨髓间充质干细胞的基础和临床研究中仍存在诸多问题需要解决。     

脂肪与骨髓来源间充质干细胞生物学特性的比较☆

背景：近几年来脂肪来源的间充质干细胞因其取材容易也被广泛研究。 目的：比较脂肪来源和骨髓来源间充质干细胞的生物学特性。 方法：分离及体外培养人骨髓源间充质干细胞和脂肪源间充质干细胞,比较它们的表型、细胞倍增时间及分泌因子水平等。 结果与结论：脂肪来源和骨髓来源的间充质干细胞在细胞表型上类似,只有CD106的表达有差异。脂肪来源间充质干细胞增殖速率比骨髓来源的间充质干细胞快。在相同体积的脂肪组织中能够得到的干细胞前体细胞的数量是骨髓的10倍以上。提示脂肪来源和骨髓来源的间充质干细胞具有相同功能,但脂肪组织是一个更有应用前景的干细胞来源。     

Gender difference in the neuroprotective effect of rat bone marrow mesenchymal cells against hypoxia-induced apoptosis of retinal ganglion cells

Bone marrow mesenchymal stem cells can reduce retinal ganglion cell death and effectively prevent vision loss. Previously, we found that during differentiation, female rhesus monkey bone marrow mesenchymal stem cells acquire a higher neurogenic potential compared with male rhesus monkey bone marrow mesenchymal stem cells. This suggests that female bone marrow mesenchymal stem cells have a stron-ger neuroprotective effect than male bone marrow mesenchymal stem cells. Here, we ifrst isolated and cultured bone marrow mesenchymal stem cells from female and male rats by density gradient centrifugation. Retinal tissue from newborn rats was prepared by enzymatic digestion to obtain primary retinal ganglion cells. Using the transwell system, retinal ganglion cells were co-cultured with bone marrow mesenchymal stem cells under hypoxia. Cell apoptosis was detected by lfow cytometry and caspase-3 activity assay. We found a marked increase in apoptotic rate and caspase-3 activity of retinal ganglion cells after 24 hours of hypoxia compared with normoxia. Moreover, apoptotic rate and caspase-3 activity of retinal ganglion cells signiifcantly decreased with both female and male bone marrow mesenchymal stem cell co-culture under hypoxia compared with culture alone, with more signiifcant effects from female bone marrow mesenchymal stem cells. Our results indicate that bone marrow mesenchymal stem cells exert a neuroprotective effect against hypoxia-induced apoptosis of retinal ganglion cells, and also that female cells have greater neuroprotective ability compared with male cells.     

Bone marrow mesenchymal stem cell therapy in ischemic stroke：mechanisms of action and treatment optimization strategies

Animal and clinical studies have conifrmed the therapeutic effect of bone marrow mesenchymal stem cells on cerebral ischemia, but their mechanisms of action remain poorly understood. Here, we summarize the transplantation approaches, directional migration, differentiation, replacement, neural circuit reconstruction, angiogenesis, neurotrophic factor secretion, apoptosis, immunomodulation, multiple mechanisms of action, and optimization strategies for bone marrow mesenchymal stem cells in the treatment of ischemic stroke. We also explore the safety of bone marrow mesenchymal stem cell transplantation and conclude that bone marrow mesenchymal stem cell transplantation is an important direction for future treatment of cerebral ischemia. Determining the optimal timing and dose for the transplantation are important directions for future research.     

骨髓基质细胞诱导分化成骨方法及相关研究进展*☆

骨髓基质细胞在体外适当的培养条件下,可以向成骨细胞、成软骨细胞、成纤维细胞、脂肪细胞、成肌细胞等多种间充质来源的中胚层组织细胞分化,取材方便、对机体损伤小,是目前最为合适的骨组织工程种子细胞。现阶段诱导骨髓基质细胞分化成骨的方法大致分为以下6种：在化学药物作用下分化、在细胞因子作用下分化、与骨细胞共培养下的分化、物理方法下的分化、中药提取物作用下的分化、转基因作用下的分化。很多诱导骨髓基质细胞分化为成骨细胞的研究结果对于深入了解诱导分化机制有重要的指导和参考价值,但仍需进一步分析各种微环境因素在诱导分化过程中的作用机制。     

Differentiation of mesenchymal stem cells into neuronal cells on fetal bovine acellular dermal matrix as a tissue engineered nerve scaffold

The purpose of this study was to assess fetal bovine acellular dermal matrix as a scaffold for supporting the differentiation of bone marrow mesenchymal stem cells into neural cells following induction with neural differentiation medium.We performed long-term,continuous observation of cell morphology,growth,differentiation,and neuronal development using several microscopy techniques in conjunction with immunohistochemistry.We examined specific neuronal proteins and Nissl bodies involved in the differentiation process in order to determine the neuronal differentiation of bone marrow mesenchymal stem cells.The results show that bone marrow mesenchymal stem cells that differentiate on fetal bovine acellular dermal matrix display neuronal morphology with unipolar and bi/multipolar neurite elongations that express neuronal-specific proteins,including βIII tubulin.The bone marrow mesenchymal stem cells grown on fetal bovine acellular dermal matrix and induced for long periods of time with neural differentiation medium differentiated into a multilayered neural network-like structure with long nerve fibers that was composed of several parallel microfibers and neuronal cells,forming a complete neural circuit with dendrite-dendrite to axon-dendrite to dendrite-axon synapses.In addition,growth cones with filopodia were observed using scanning electron microscopy.Paraffin sectioning showed differentiated bone marrow mesenchymal stem cells with the typical features of neuronal phenotype,such as a large,round nucleus and a cytoplasm full of Nissl bodies.The data suggest that the biological scaffold fetal bovine acellular dermal matrix is capable of supporting human bone marrow mesenchymal stem cell differentiation into functional neurons and the subsequent formation of tissue engineered nerve.     

Chitosan-collagen porous scaffold and bone marrow mesenchymal stem cell transplantation for ischemic stroke

In this study, we successfully constructed a composite of bone marrow mesenchymal stem cells and a chitosan-collagen scaffold in vitro, transplanted either the composite or bone marrow mesenchymal stem cells alone into the ischemic area in animal models, and compared their effects. At 14 days after co-transplantation of bone marrow mesenchymal stem cells and the hitosan-collagen scaffold, neurological function recovered noticeably. Vascular endothelial growth factor expression and nestin-labeled neural precursor cells were detected in the ischemic area, surrounding tissue, hippocampal dentate gyrus and subventricular zone. Simultaneously, a high level of expression of glial fibrillary acidic protein and a low level of expression of neuron-specific enolase were visible in Brd U-labeled bone marrow mesenchymal stem cells. These findings suggest that transplantation of a composite of bone marrow mesenchymal stem cells and a chitosan-collagen scaffold has a neuroprotective effect following ischemic stroke.     

Collagen-chitosan scaffold impregnated with bone marrow mesenchymal stem cells for treatment of traumatic brain injury

Combinations of biomaterials and cells can effectively target delivery of cells or other therapeutic factors to the brain to rebuild damaged nerve pathways after brain injury.Porous collagen-chitosan scaffolds were prepared by a freeze-drying method based on brain tissue engineering.The scaffolds were impregnated with rat bone marrow mesenchymal stem cells.A traumatic brain injury rat model was established using the 300 g weight free fall impact method.Bone marrow mesenchymal stem cells/collagen-chitosan scaffolds were implanted into the injured brain.Modified neurological severity scores were used to assess the recovery of neurological function.The Morris water maze was employed to determine spatial learning and memory abilities.Hematoxylin-eosin staining was performed to measure pathological changes in brain tissue.Immunohistochemistry was performed for vascular endothelial growth factor and for 5-bromo-2-deoxyuridine(BrdU)/neuron specific enolase and BrdU/glial fibrillary acidic protein.Our results demonstrated that the transplantation of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds to traumatic brain injury rats remarkably reduced modified neurological severity scores,shortened the average latency of the Morris water maze,increased the number of platform crossings,diminished the degeneration of damaged brain tissue,and increased the positive reaction of vascular endothelial growth factor in the transplantation and surrounding areas.At 14 days after transplantation,increased BrdU/glial fibrillary acidic protein expression and decreased BrdU/neuron specific enolase expression were observed in bone marrow mesenchymal stem cells in the injured area.The therapeutic effect of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds was superior to stereotactic injection of bone marrow mesenchymal stem cells alone.To test the biocompatibility and immunogenicity of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds,immunosuppressive cyclosporine was intravenously injected 12 hours before transplantation and 1-5 days after transplantation.The above indicators were similar to those of rats treated with bone marrow mesenchymal stem cells and collagen-chitosan scaffolds only.These findings indicate that transplantation of bone marrow mesenchymal stem cells in a collagen-chitosan scaffold can promote the recovery of neuropathological injury in rats with traumatic brain injury.This approach has the potential to be developed as a treatment for traumatic brain injury in humans.All experimental procedures were approved by the Institutional Animal Investigation Committee of Capital Medical University,China(approval No.AEEI-2015-035)in December 2015.     

骨髓间充质干细胞防治心肌细胞凋亡的基因及蛋白研究*★

背景：骨髓间充质干细胞移植后可作用于心肌细胞，防止心肌细胞损伤的发生或继发性的病变。 目的：分析骨髓间充质干细胞对心肌细胞凋亡的基因蛋白表达影响。 方法：由第一作者检索1997/2010 PubMed数据及万方数据库有关骨髓间充质干细胞、心肌细胞凋亡以及基因蛋白表达等方面的文献。 结果与结论：骨髓间充质干细胞以其获取方便、分化能力强、低免疫原性、无排斥反应等特点成为研究的重点。骨髓间充质干细胞能够作用于心肌细胞，防止心肌细胞损伤的发生或继发性的病变，主要是通过调控Bcl-2、Bax、Fas、FasL、Caspase等基因蛋白的表达，抑制心肌细胞凋亡的发生，对于运动过程中出现的因心肌细胞凋亡而影响其成绩下降的现象具有一定的保护作用和应用价值。     

三维支架中骨髓间充质干细胞及软骨细胞和脂肪源性成熟基质细胞的成软骨效应

背景：软骨修复的关键是种子细胞在三维支架中的定向分化,但细胞放入三维支架中很难有透明软骨表现。 目的：考察3种种子骨髓间充质干细胞,软骨细胞和脂肪源性成熟基质细胞在支架中的成软骨特性。 方法：抽取羊骨髓,胰酶消化后获得原代骨髓间充质干细胞细胞、软骨细胞和脂肪源性成熟基质细胞,单层培养扩增。取P1骨髓间充质干细胞,P3软骨细胞和P3脂肪源性成熟基质细胞种植入不同比例(10%,20%,50%,80%,100%)的胶原/透明质酸支架中, 在无血清培养液中三维培养。2周后,用SO染色和免疫组织化学染色分析,观察硫酸软骨素和Ⅱ型胶原合成情况。 结果与结论：种子细胞在三维支架中特定条件下有成软骨效应,骨髓间充质干细胞在含20%透明质酸的胶原/透明质酸支架扩增少于3代有成软骨效应,软骨细胞传代数更高仍然有成软骨效应,但脂肪源性成熟基质细胞成软骨效应能力较弱。结果提示,在同样条件下骨髓间充质干细胞、软骨细胞较脂肪源性成熟基质细胞有更好的成软骨性能。 关键词：骨髓间充质干细胞;软骨细胞;组织工程;软骨;脂肪干细胞 doi:10.3969/j.issn.1673-8225.2010.16.004     

Dorsal root ganglion neurons promote proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells

Preliminary animal experiments have confirmed that sensory nerve fibers promote osteoblast differentiation, but motor nerve fibers have no promotion effect. Whether sensory neurons promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells remains unclear. No results at the cellular level have been reported. In this study, dorsal root ganglion neurons(sensory neurons) from Sprague-Dawley fetal rats were co-cultured with bone marrow mesenchymal stem cells transfected with green fluorescent protein 3 weeks after osteogenic differentiation in vitro, while osteoblasts derived from bone marrow mesenchymal stem cells served as the control group. The rat dorsal root ganglion neurons promoted the proliferation of bone marrow mesenchymal stem cell-derived osteoblasts at 3 and 5 days of co-culture, as observed by fluorescence microscopy. The levels of m RNAs for osteogenic differentiation-related factors(including alkaline phosphatase, osteocalcin, osteopontin and bone morphogenetic protein 2) in the co-culture group were higher than those in the control group, as detected by real-time quantitative PCR. Our findings indicate that dorsal root ganglion neurons promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, which provides a theoretical basis for in vitro experiments aimed at constructing tissue-engineered bone.