Shear stress and VEGF enhance endothelial differentiation of human adipose-derived stem cells

Abstract

Herein we combine chemical and mechanical stimulation to investigate the effects of vascular endothelial growth factor (VEGF) and physiological shear stress in promoting the differentiation human adipose derived stem cells (ADSCs) into endothelial cells. ADSCs were isolated and characterized; endothelial differentiation was promoted by culturing confluent cells in 50?ng/ml VEGF under physiological shear stress for up to 14 days. Afterwards, endothelial cells were seeded onto collagen or acellular aortic valve matrices and exposed to four culture conditions: shear stress + VEGF; shear stress ? VEGF; static + VEGF and static ? VEGF. After 7 days, phenotype was investigated. ADSCs subjected to shear stress and VEGF express a comprehensive range of specific endothelial markers (vWF, eNOS and FLT-1 after 7 days and CD31, FLk-1 and VE-cadherin after 14 days) and maintain the phenotype when seeded onto scaffolds. Our protocol proved to be an efficient source of endothelial-like cells for tissue engineering based on autologous ADSC.     

VEGF and BMP-6 enhance bone formation mediated by cloned mouse osteoprogenitor cells

New strategies such as combined utilization of growth factors may provide a better treatment for difficult fractures. We have demonstrated enhanced angiogenesis and osteogenesis through the actions of vascular endothelial growth factor (VEGF) and bone morphogenetic protein-6 (BMP-6) on the osteogenic differentiation of a cloned mouse osteoprogenitor cell in vitro and ectopic bone formation in vivo. Human VEGF and BMP-6 genes expressed together produced a significant increase in alkaline phosphatase activity, expression of the RunX2 and osteocalcin genes and mineralization. Microcomputed tomographic analysis of subcutaneous implants consisting of cells transfected with VEGF and BMP-6 cDNA and delivered on a 3D poly (lactic-co-glycolic acid) scaffold confirmed the additive effects between VEGF and BMP-6. Ectopic bone formation in the VEGF plus BMP-6 group was greatest compared to that in either VEGF or BMP-6 alone. This is the first study that demonstrates osteogenesis in vitro and in vivo through the additive effects of VEGF and BMP-6.     

骨髓间充质干细胞成骨分化调控的研究进展

骨髓间充质干细胞具有较强的自我更新能力和多向分化潜能 ,其应用是目前国际上组织工程领域中重要的研究内容之一。近年来 ,许多实验室从分子、生化、物理等水平对其成骨分化调控进行了深入研究 ,取得了较大的进展     

Collagen I gel promotes homogenous osteogenic differentiation of adipose tissue-derived mesenchymal stem cells in serum-derived albumin scaffold

Repair of bone defects is a difficult clinical problem for reconstructive surgeons. Bone tissue engineering using an appropriate scaffold with cells is a new therapy for the repair of bone defects. The aim of this study was to evaluate the in vitro osteogenesis of canine adipose tissue-derived mesenchymal stem cells (Ad-MSCs) cultured in a combination of collagen I gel and a porous serum-derived albumin scaffold. A serum-derived albumin scaffold was prepared with canine serum by cross-linking and freeze-drying procedures. Ad-MSCs were seeded into serum-derived albumin scaffolds with or without collagen I gel, and were exposed to osteogenic differentiation conditions in vitro. After 28?days of in vitro culture, the distribution and osteogenic differentiation of Ad-MSCs cultured in the scaffold were evaluated by scanning electron microscopy, histology, immunohistochemistry, alkaline phosphatase (ALP) activity assay, and calcium colorimetric assay. Ad-MSCs showed more homogeneous distribution and osteogenic differentiation in the scaffold with collagen I gel than without collagen I gel. ALP activity and extracellular matrix mineralization in the construct with type I collagen were significantly higher than in the construct without type I collagen (p?<?0.05). In conclusion, the combination of collagen I gel and the serum-derived albumin scaffold enhanced osteogenic differentiation and homogenous distribution of Ad-MSCs.     

BMP-4 increases activin A gene expression during osteogenic differentiation of mouse embryonic stem cells

Abstract

Activin A is a growth factor released by mature osteoblasts that has a critical effect on bone formation. We investigated the effect of bone morphogenetic protein (BMP)-4 on activin A gene expression during in vitro osteogenic differentiation of mouse embryonic stem (ES) cells. Embryoid bodies were cultured in retinoic acid (RA) for three days and then without RA for two days. Seeded cells received osteogenic medium with β-glycerophosphate, L-ascorbic acid 2-phosphate and dexamethasone during 19 days, with or without BMP-4. Six independent experiments were carried out. Real-time PCR was used to detect gene expression of activin A, Oct-4, Nanog, osteocalcin, RUNX2 and bone alkaline phosphatase. Immunofluorescence was used to co-localize activin A with the undifferentiation marker stage-specific embryonic antigen 1. Cells treated with BMP-4 had an increased gene expression of activin A, osteocalcin and bone alkaline phosphatase (p?<?0.05). In conclusion, BMP-4 increases activin A gene expression during mouse ES cell differentiation into bone precursors.     

Osteogenic response of human adipose-derived stem cells to BMP-6, VEGF,and combined VEGF plus BMP-6 in vitro

Exogenous addition of three factors—mesenchymal stem cells (MSCs), vascular endothelial growth factor (VEGF), and bone morphogenetic proteins (BMPs)—has proven to be more beneficial than delivery of any single factor for fracture repair in animal models. We studied the osteogenic differentiation of human adipose-derived stem cells (hADSCs) in the presence of VEGF, BMP-6, or VEGF plus BMP-6 to better understand their enhancement of osteoblastic differentiation of MSCs. The VEGF plus BMP-6 group demonstrated an additive effect on the enhancement of mineralization and expression of ALP and Msx2 genes. Unlike VEGF or BMP-6 alone, the combination of VEGF and BMP-6 significantly enhanced the expression of COL1A1, osterix, and Dlx5 genes. The data indicate that a cross-talk between VEGF and BMP-6 signaling pathways enhances osteogenic differentiation of hADSCs.     

Effect of the fiber diameter and porosity of non-woven PET fabrics on the osteogenic differentiation of mesenchymal stem cells

The proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs) was investigated in three-dimensional non-woven fabrics prepared from polyethylene terephthalate (PET) fiber with different diameters. When seeded into the fabrics of cell scaffold, more MSC attached in the fabric of thicker PET fibers than that of thinner ones, irrespective of the fabric porosity. The morphology of cells attached became more spreaded with an increase in the fiber diameter of fabrics. The rate of MSC proliferation depended on the PET fiber diameter and porosity of fabrics: the bigger the fiber diameter of fabrics with higher porosity, the higher their proliferation rate. When the alkaline phosphatase (ALP) activity and osteocalcin content of MSC cultured in different types of fabrics was measured to evaluate the ostegenic differentiation, they became maximum for the non-woven fabrics with a fiber diameter of 9.0 μm, although the values of low-porous fabrics were significantly high compared with those of high porous fabrics. We concluded that the attachment, proliferation and bone differentiation of MSC was influenced by the fiber diameter and porosity of non-woven fabrics as the scaffold.     

不同年龄段大鼠脂肪间充质干细胞增殖及成骨能力的研究

通过研究年龄对大鼠脂肪间充质干细胞增殖、成骨分化的影响,从而为临床寻找理想的种子细胞来源。用密度梯度离心法分离不同年龄段脂肪间充质干细胞进行培养,并保留贴壁细胞传代,观察细胞生长情况,检测其增殖活性,诱导后茜素红染色,钙离子浓度测定。结果显示,传代细胞的增殖速度比原代细胞快,但随着年龄的增长脂肪间充质干细胞的增殖能力下降。诱导条件下,各年龄组均出现矿化结节,但钙离子浓度随年龄增加而降低。本实验提示,脂肪间充质干细胞增殖和成骨分化能力随着年龄的增加而降低,但各年龄段脂肪间充质干细胞经过体外增殖、诱导后均可满足临床应用中不同患者的要求。     

补骨脂素对大鼠骨髓间充质干细胞成骨分化的影响

目的研究不同浓度的补骨脂素对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)成骨分化的影响,为临床应用补骨脂素治疗骨质疏松症提供理论依据。方法选取8周龄健康雄性SD大鼠,取双侧股骨、胫骨,对大鼠的BMMSCs进行分离、原代培养和细胞表型鉴定。取第3代大鼠BMMSCs,体外利用成骨诱导培养基进行不同浓度补骨脂素的药物诱导,MTT法检测不同浓度的补骨脂素对大鼠BMMSCs生长的影响;通过碱性磷酸酶(ALP)活性测定、Western blot法和茜素红染色方法评价不同浓度补骨脂素对大鼠BMMSCs成骨分化能力。结果第3代大鼠BMMSCs表面抗原符合干细胞鉴定标准,成骨诱导后给予15μmol/L补骨脂素对大鼠BMMSCs的增殖作用最佳,ALP活性、Runx-2表达和钙化结节数目均明显高于经典成骨诱导组、5μmol/L和10μmol/L和20μmol/L补骨脂素组。结论15μmol/L补骨脂素成骨诱导促进BMMSCs向成骨分化,从而起到防治骨质疏松的作用。     

Heterodimeric BMP-2/7 exhibits different osteoinductive effects in human and murine cells

As robust osteoinductive cytokines, bone morphogenetic proteins (BMPs) play a significant role in bone tissue engineering. Constituted of two different polypeptides, heterodimeric BMPs are more effective than the homodimers in bone formation. While most studies focused on the murine cell lines, such as murine preosteoblasts MC3T3-E1, the role of heterodimeric BMPs in the osteogenic differentiation of human cells remains uncertain, which hinders their application to practical treatment. In this study, we compared the osteoinductive effects of BMP-2/7 heterodimer in human adipose-derived stem cells (hASCs) with their homodimers BMP-2 and BMP-7, in which MC3T3-E1 cells were utilized as a positive control. The results indicated that BMP-2/7 was not a stronger inducer during the osteogenic differentiation of hASCs as that for MC3T3-E1, and extracellular-signal-regulated kinase signaling played a role in the different effects of BMP-2/7 between hASCs and MC3T3-E1. Our study demonstrates the osteoinductive effects of heterodimeric BMP-2/7 present in a cell-specific pattern and cautions should be taken when applying heterodimeric BMP-2/7 to clinical practice.     

Continuous stimulation with differentiation factors is necessary to enhance osteogenic differentiation of human mesenchymal stem cells in-vitro

Bone defect treatment belongs to the most challenging fields in orthopedic surgery and requires the well-coordinated application of mesenchymal stem cells (MSC) and differentiation factors. MSC isolated from reaming material (RMSC) and iliac crest (BMSC) in combination with bone morphogenetic protein-7 (BMP-7) and insulin-like growth factor-1 (IGF-1) have been used. The short half-life of both factors limit their applications: a burst release of the factor can probably not induce sustainable differentiation. We stimulated MSC in osteogenic differentiation medium with three different concentrations of BMP-7 or IGF-1: Group A was stimulated continuously, group B for 24?h and group C remained without any stimulation. Osteogenic differentiation was measured after seven and 14?days by alizarin red staining and alkaline phosphatase (ALP) activity. Continuous stimulation led to higher levels of osteogenic differentiation than short-term stimulation. This could lead to a reconsideration of established application forms for differentiation factors, aiming to provide a more sustained release.     

长链非编码RNA AK089560在间质干细胞成骨与成脂分化中的表达

背景：最近研究发现众多长链非编码RNA调控干细胞多潜能性和分化。长链非编码RNA AK089560在干细胞多向分化中的表达变化及作用,目前尚不清楚。目的：观察间质干细胞C3H10T1/2成骨分化与成脂分化过程中AK089560的表达。方法：重组人骨形态发生蛋白2诱导间质干细胞C3H10T1/2成骨分化,碱性磷酸酶染色鉴定早期成骨分化。地塞米松、吲哚美辛和胰岛素三因子联合诱导C3H10T1/2成脂分化,油红O染色鉴定成脂分化结果。采用qRT-PCR检测诱导前后不同时间点AK089560表达的变化。采用RNAfold软件预测AK089560二级结构,UCSC基因组浏览器分析AK089560邻近编码蛋白基因,fancyGENE在线软件构件长链非编码RNA与编码基因关系图。结果与结论：C3H10T1/2经成骨诱导后,70%以上细胞碱性磷酸酶染色阳性;成脂分化诱导后,80%以上细胞油红O染色阳性。qRT-PCR结果显示,长链非编码RNA AK089560在成骨分化与成脂分化第2,4,6天表达均明显下降,成骨分化与成脂分化第2,4,6天与相应时间未分化组相比均差异有显著性意义(P < 0.05)。生物信息学分析表明,LncRNA AK089560具有复杂茎环结构,与编码基因Sema3a形成sense overlap关系。结果表明AK089560在间质干细胞成骨与成脂分化中下调表达,提示其可能参与调控间质干细胞多向分化。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;组织工程全文链接：     

成骨诱导因子Nell-1及Noggin短发卡RNA联合转染脂肪间充质干细胞的促成骨分化能力

背景:Nell-1在诱导成骨分化和新骨形成中的作用已有报道,但试图通过双基因联合转染起协同成骨作用的研究很少。目的:体外环境下抑制细胞中Noggin基因的同时上调Nell-1基因,观察其对脂肪间充质干细胞成骨分化能力的影响。方法:取健康成年大鼠脂肪组织获取脂肪间充质干细胞。将细胞分为3组,对照组转染空载体病毒Lv-EGFP(增强型绿色荧光蛋白),Nell-1组单纯转染Lv-Nell-1,Nell-1+Noggin shRNA组同时转染Lv-Nell-1和Lv-Noggin shRNA。转染后3,7,14 d,应用实时荧光定量PCR检测Nell-1及成骨相关标志物(Col-Ⅰ、ALP、OCN)的表达,转染后14 d行茜素红染色。结果与结论:①转染后3 d,对照组Nell-1 mRNA相对表达量与Nell-1组比较,差异无显著性意义(P>0.05)。与Nell-1+Noggin shRNA组比较,对照组和Nell-1组Nell-1 mRNA表达量偏低,差异均有显著性意义(P<0.05)。转染后7,14 d,Nell-1+Noggin shRNA组Nell-1 mRNA相对表达量仍高于对照组和Nell-1组,且Nell-1组高于对照组,两两比较差异均有显著性意义(P<0.05);②转染后3,7,14 d,Nell-1+Noggin shRNA组各成骨基因mRNA相对表达量均显著高于对照组和Nell-1组,且Nell-1组高于对照组,组间比较差异有显著性意义(P<0.05);③转染后14 d,Nell-1+Noggin shRNA组呈现较多细胞聚集形成的结节,Nell-1组钙结节数量少于Nell-1+Noggin shRNA组,对照组未见明显被染色的钙结节;④实验结果表明,Nell-1基因可以促进大鼠脂肪间充质干细胞成骨分化,而干扰Noggin基因会上调Nell-1基因的表达,形成显著的协同成骨分化作用。     

体外构建人羊膜间充质干细胞膜片及成骨分化潜能

文题释义：细胞膜片技术：该技术避免了蛋白酶的消化和外源性支架材料的应用,通过细胞外基质分泌形成膜片组织,然后将膜片用于修复组织缺损和改善器官功能。该技术保留了大量自体细胞分泌的细胞外基质,为细胞的增殖和分化提供与体内极度相似的微环境,目前该技术已经用于临床眼角膜和食管损伤的修复。人羊膜间充质干细胞：取自于废弃的胎盘,贴壁生长,具有低免疫原性和生长周期短等特点,不仅具有成体间充质干细胞的特性还具有部分胚胎间充质干细胞的特性。  摘要背景：人羊膜间充质干细胞属于成体干细胞,其来自于废弃的胎盘,来源广泛,可以无创获取,具有免疫原性低、生长周期短等特点,是组织工程种子细胞的重要来源,目前人羊膜间充质干细胞已经用于临床糖尿病的治疗。目的：探索一种简便的方法构建人羊膜间充质干细胞膜片,并研究其成骨分化潜能。方法：将第3代人羊膜间充质干细胞高密度接种于普通培养皿中,加入成膜片诱导培养基以构建人羊膜间充质干细胞膜片,通过组织学染色以及扫描电镜观察细胞膜片的特性。取第3代人羊膜间充质干细胞高密度接种于培养皿中,加入成膜片诱导培养基培养7 d,再换用成骨诱导培养基培养14 d以构建成骨诱导的人羊膜间充质干细胞膜片。通过茜素红染色、免疫组化染色、碱性磷酸酶活性、RT-PCR以及Western blot检测人羊膜间充质干细胞膜片的成骨分化潜能。结果与结论：①苏木精-伊红染色可见人羊膜间充质干细胞膜片由多层细胞累积而成,细胞分布均匀;②扫描电镜观察可见人羊膜间充质干细胞膜片呈复层结构,胞外有大量的胞外基质产生,细胞包埋于胞外基质中;③人羊膜间充质干细胞膜片成骨诱导14 d,茜素红染色后可见橘红色沉淀,免疫组化染色后细胞周围有大量Ⅰ型胶原产生;④与未诱导的人羊膜间充质干细胞膜片相比,成骨诱导14 d后人羊膜间充质干细胞膜片碱性磷酸酶活性显著升高(P < 0.01),Ⅰ型胶原、骨钙蛋白、Runt相关转录子2的mRNA和蛋白表达量显著升高(P < 0.05);⑤该实验应用一种简便、经济的方法在普通培养皿上成功构建了人羊膜间充质干细胞膜片,体外研究证实人羊膜间充质干细胞膜片具有良好的成骨分化潜能。ORCID: 0000-0003-2163-3897(邹刚)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     

辛伐他汀可刺激骨髓间充质干细胞的成骨分化

背景:辛伐他汀可显著刺激骨髓间充质干细胞成骨分化,但机制不明.近年研究证实p38 MAPK信号通路参与调控骨髓间充质干细胞向成骨细胞分化的过程.目的:观察p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路在辛伐他汀体外干预刺激骨髓间充质干细胞成骨分化中的作用....     

人生长激素对人牙周膜干细胞增殖及成骨分化的影响

背景:生长激素已被证明对于非牙源性间充质干细胞的增殖和骨向分化有促进作用,但对于人牙周膜干细胞的生物学效应还不明确.目的:探讨人生长激素对人牙周膜干细胞增殖以及成骨分化的影响.方法:用含0(对照组),10,100,200μg/L人生长激素的α-MEM完全培养基干预第3代人牙周膜干细胞,分别在第1,3,5,7天采用CCK...     

miR-146a调控骨髓间充质干细胞成骨分化的机制研究

目的探讨miR-146a调控骨髓间充质干细胞(BMSC)成骨分化的作用及其分子机制。方法贴壁法分离培养小鼠BMSC,检测成骨分化早期标志物Runx 2的变化,观察BMSC体外成骨分化,利用miRNA特异性的聚合酶链式反应(miRNAspecific qPCR)观察miR-146a的变化情况,并干预miR-146a表达,明确miR-146a对BMSC成骨分化的调控作用。结果成功建立了稳定的BMSC体外培养体系,该细胞能够成功分化为脂肪细胞和成骨细胞;在成骨诱导培养条件下,随着成骨分化,miR-146a水平降低,过表达miR-146a,成骨分化早期标志分子Runx 2表达降低;转染miR-146a拮抗体antago-miR-146a可以补救Runx 2表达的降低。结论 miR-146a负向调控BMSC成骨分化,拮抗miR-146a可以补救BMSC成骨分化的降低。     

锌离子浓度可影响兔骨髓间充质干细胞的增殖与成骨分化#br#

文题释义：骨髓间充质干细胞：间充质干细胞是一种多能干细胞,最早于骨髓中发现,后来陆续在外周血、脂肪等组织中被分离出来。相比于其他来源的干细胞,骨髓间充质干细胞不仅具有自我更新及多向分化潜能,同时还能分泌某些有助于血管生成的细胞因子;在骨修复方面,其能在促成骨的同时促进局部毛细血管的再生,有较好的应用前景。 成骨分化：人体内干细胞在生长增殖过程中,受各种生物因素如细胞调控因子、激素水平以及细胞微环境中微量元素的影响,可向不同组织分化。影响成骨分化的因素包括降钙素、锌离子、铜离子、钙离子、骨形态发生蛋白2、生长分化因子5等,因此量化每一个影响因素对于骨组织工程具有重要意义。 背景：锌离子是人体所必需的金属微量元素,在抑制破骨细胞活性和促进细胞成骨分化方面起着重要作用。然而,不同浓度锌离子对兔骨髓间充质干细胞增殖及成骨分化的影响却鲜有研究。 目的：评价不同浓度锌离子对兔骨髓间充质干细胞增殖与成骨分化的影响,探索最适宜的锌离子浓度。   方法：从乳兔长骨骨髓内提取骨髓间充质干细胞培养并传代,分为6组,即10^-4,10^-5,10^-6,10^-7,10^-8 mol/L锌离子组以及空白对照组,采用CCK-8法检测细胞增殖活性并绘制生长曲线,碱性磷酸酶试剂盒评价细胞的成骨分化能力,活/死细胞染色表征细胞的生长活性,茜素红染色观察细胞的矿化能力,荧光定量PCR检测成骨相关基因的表达。   结果与结论：①除10^-4 mol/L锌离子组之外,随着时间延长,各组细胞数量呈增长趋势,且10^-7 mol/L锌离子组在培养5 d后细胞数量最多,而10^-4 mol/L锌离子组细胞数量最少;②细胞培养14 d后,当锌离子浓度为10^-7 mol/L时,碱性磷酸酶活性最高,10^-4 mol/L时碱性磷酸酶表达量最低;③活/死细胞染色显示10^-7 mol/L锌离子组活细胞数量最多,10^-4 mol/L锌离子组则几乎全为死细胞;④细胞培养14 d后,当锌离子浓度为   10^-7 mol/L时,可观察到明显的钙结节形成,且成骨相关基因表达水平最高;⑤结果表明,锌离子在一定浓度范围内,能有效促进兔骨髓间充质干细胞的增殖与成骨分化,且浓度为10^-7 mol/L时,其促增殖、成骨诱导及促矿化能力最强。因此,在培养基中添加适宜浓度锌离子,有利于兔骨髓间充质干细胞的成骨分化。 ORCID: 0000-0003-1353-2148(赵桥) 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     

全骨髓贴壁法培养兔骨髓间充质干细胞体外定向成骨诱导分化及鉴定

背景：流式细胞仪分离法和免疫磁珠分离法对细胞活性影响较大,密度梯度离心法虽然能够获得纯度高的单核细胞,但由于多次离心可造成细胞的大量流失且对细胞活性有一定的影响使其应用值得商榷。 目的：采用全骨髓贴壁法分离兔骨髓间充质干细胞进行成骨诱导分化及鉴定。 方法：采用全骨髓贴壁法体外分离培养兔骨髓间充质干细胞,倒置显微镜下观察细胞形态学特征。在成骨诱导剂作用下,通过碱性磷酸酶染色试剂盒行碱性磷酸酶染色,Ⅰ型胶原免疫细胞化学染色,Von Kossa法及茜素红进行矿化结节染色以及电镜下检测兔骨髓间充质干细胞成骨诱导后的形态结构。 结果与结论：经诱导后细胞出现与成骨细胞相似的形态学特征,碱性磷酸酶染色阳性,Ⅰ型胶原免疫细胞化学染色,Von-Kossa法及茜素红矿化结节染色阳性。表明经成骨诱导剂诱导后全骨髓贴壁法体外分离纯化培养的兔骨髓间充质干细胞能向成骨细胞方向分化增殖。     

人骨髓间充质干细胞体外成骨与成脂分化过程中FGF1及其受体表达的变化

目的 研究人骨髓间充质干细胞(hBMSCs)成骨与成脂分化过程中成纤维细胞生长因子1(FGF1)及其受体(FGFRs)的动态表达,探讨FGF1在hBMSCs成骨与成脂分化过程中的作用.方法 密度梯度离心法分离hBMSCs,取第3代细胞分别进行成骨和成脂诱导分化并染色鉴定.通过real-time PCR方法检测成骨和成脂标志基因以及FGF1/FGFRs的表达,免疫荧光染色法检测诱导分化前后FGF1的表达.Real-time PCR及油红O染色鉴定外源性FGF1对hBMSCs成骨与成脂分化的影响.结果 体外诱导hBMSCs成骨和成脂分化后茜素红和油红O染色结果呈阳性;FGF1和FGFRs在诱导过程中呈动态表达;免疫荧光染色结果表明FGF1主要在细胞质中分布,成骨诱导3d后在细胞核中的表达增加;相比对照组,加入外源的FGF1组7d后脂滴数量明显减少,成脂标志基因表达受到抑制(P＜0.01),而加入外源的FGF1 3 d骨桥蛋白(OPN)表达升高(P＜0.05).结论 FGF1能够抑制hBMSCs的成脂分化,并且可能通过促进骨桥蛋白表达而提前hBMSCs的成骨矿化.