碱性成纤维细胞生长因子重组腺病毒转染兔骨髓间充质干细胞的表型变化

碱性成纤维细胞生长因子对体外培养的兔骨髓基质细胞生长特性的影响王志勇;陈增海;于胜吉【期刊名称】《山东大学学报：医学版》【年(卷),期】2006(44)11【摘要】目的:研究骨髓基质细胞(BMSC)在体外培养条件下分化为成骨细胞的能力,及碱性成纤维细胞生长因子(bFGF)对其贴附、增殖、分化的影响。

方法:将各组BMSC传代后加入bFGF,其浓度分别为0、5、10、50、1002、00 ng/ml,用倒置相差显微镜、电镜、四唑盐比色法、碱性磷酸酶(ALP)染色、钙染色和电子探针等方法观察其形态、贴附、生长增殖活性、分化成熟和体外钙化的状况。

结果:经10 ng/ml bFGF诱导后,BMSC贴附细胞数量明显增加。

当100 ng/ml时,bFGF对BMSC的促增殖作用达到最大值。

当加入bFGF时,ALP染色阳性率降低,当bFGF 浓度为100μg/L时阳性率最低(P<0.01)。

传代细胞连续培养30 d后钙染色结果呈阳性,电子探针检测矿化区的Ca/P值与羟基磷灰石结晶中Ca/P值相符。

电镜观察可见培养的细胞具有典型的成骨细胞形态特征,并能产生胶原纤维。

结论:BMSC 在体外培养条件下具有分化为成骨细胞的能力,bFGF能够促进细胞的贴壁和增殖,但对其分化成熟具有抑制作用。

【总页数】5页(P1090-1094)【关键词】骨髓基质细胞;成骨细胞;成纤维细胞生长因子2;兔【作者】王志勇;陈增海;于胜吉【作者单位】山东大学齐鲁医院急诊科;山东大学第二医院骨科;中国医学科学院肿瘤医院骨科【正文语种】中文【中图分类】Q954.658【相关文献】1.不同浓度碱性成纤维细胞生长因子对兔骨髓基质细胞的影响 [J], 韦永中;范卫民;陈哲峰;崔维顶2.重组人骨形态发生蛋白2与碱性成纤维细胞生长因子联合促进兔骨髓基质细胞血管内皮细胞生长因子表达及其成骨潜能 [J], 常祺;黄昌林;黄涛3.兔骨髓基质细胞与陶瓷样异种骨的体外培养实验兔骨髓基质细胞形态、生长、附着及增殖性能观察 [J], 李彦林;韩睿;黄河;李世和;曾才铭;王宏邦4.碱性成纤维细胞生长因子对兔骨髓基质细胞生物行为的影响 [J], 刘建;常祺;胡蕴玉;孟国林5.复合骨形态发生蛋白2及碱性成纤维细胞生长因子注射型骨修复材料对兔骨髓基质细胞增殖和超微结构的影响 [J], 崔赓;胡蕴玉;雷伟;孙明林;李洁;靳小兵;汪培铭;杨柳;吕昌伟;吕荣因版权原因，仅展示原文概要，查看原文内容请购买。

重组人碱性成纤维细胞生长因子对兔视网膜缺血再灌注损伤的保护作用邵宏超;葛嫣然;李利艳;王林洪;马建英;曹凤英;田华【期刊名称】《临床误诊误治》【年(卷),期】2014(000)006【摘要】目的探讨重组人碱性成纤维细胞生长因子( recombinant human basic fibroblast growth factor, rh-bFGF)对兔视网膜缺血再灌注损伤( retinal ischemia reperfusion injury, RIRI)的保护作用。

方法选取健康纯种大耳白兔66只,随机分为假手术组6只,RIRI组30只,rh-bFGF组30只。

通过升高眼压的方法制作兔RIRI模型,RIRI组予0.9%氯化钠注射液,rh-bFGF组予rh-bFGF治疗,假手术组仅用5号头皮针从耳前方角巩膜缘内刺入前房,之后缓慢拔除针头。

RIRI组和rh-bFGF组在RIRI后6 h、12 h、24 h、48 h、72 h常规HE染色观察视网膜组织形态变化,SABC免疫组织化学法检测视网膜组织中凋亡抑制基因Bcl-2和促凋亡基因Bax蛋白表达及Bcl-2/Bax的变化。

结果假手术组视网膜未见凋亡细胞；RIRI组视网膜凋亡细胞主要出现在内核层和神经节细胞层,且在24 h达到高峰；rh-bFGF组与RIRI组比较,观察各时间点细胞损伤情况基本相似。

Bcl-2和Bax在正常视网膜组织中几乎不表达,在RIRI后6 h开始表达,24 h达到高峰,48 h开始下降,72 h后表达明显减弱。

rh-bFGF组与RIRI组比较,Bcl-2表达明显增强,而Bax 表达减弱(P均<0.05)。

结论 Bcl-2和Bax参与了RIRI的调控,rh-bFGF可通过上调Bcl-2表达及下调Bax表达,使Bcl-2/Bax比值升高,起到保护视网膜组织的作用。

%Objective To explore the protective effect of recombinant human basic fibroblast growth factor (rh-bFGF) on retinal ischemia-reperfusioninjury (RIRI) in rabbits. Methods The 66 healthy purebred rabbits were randomly divided into sham-operation group (n=6), ischemia-reperfusion model group (n=30) and rh-bFGF treatment group (n=30). The RIRI models were established by elevating intraocular pressure, the ischemia-reperfusion model group was injected with 0. 9% sodium chloride injec-tion;rh-bFGF drug therapy was performed in rh-bFGF treatment group;the sham-operation group only received the 5th scalp heedle from ear piercing the anterior chamber angle in front of the sclera edge, and then the needle was slowly removed. Retina histopatho-logical changes were observed by routine hematoxylin and eosin stain 6 h, 12 h, 24 h, 48 h and 72 h after RIRI in ischemia-reperfu-sion model and rh-bFGF treatment groups. Expressions of inhibiting gene Bcl-2 and stimulative gene Bax and changes of Bax/Bcl-2 in retinal tissues were detected with SABC immunohistochemical method. Results No apoptotic cell was found in sham-operation group;apoptotic cells mainly existed in the inner nuclear layer and the ganglion cell layer, and apoptotic cells reached the peak 24 h after reperfusion;the rh-bFGF treatment groups had the similar condition with the ischemia-reperfusion model group in cell in-jury at each observational time. No expressions of Bcl-2 and Bax positive cells were found in normal retina tissues;the expressions were found 6 h after reperfusion, and the expressions reached the peak 24 h after reperfusion, and began to descented 48 h after reperfusion, and the decrease was obvious 72 h after reperfusion. Bcl-2 expression was significantly stronger, while Bax expression was significantly reduced in rh-bFGF treatment group,compared with those in ischemia-reperfusion model group (P<0. 05). Con-clusion Bcl-2 and Bax protein are involved in the RIRI regulation. The retina tissue can be protected with Bcl-2 expression of up-regulation and the Bax expression of down-regulation by rh-bFGF, which can increase the Bcl-2/Bax ratio.【总页数】5页(P100-104)【作者】邵宏超;葛嫣然;李利艳;王林洪;马建英;曹凤英;田华【作者单位】063000 河北唐山，河北联合大学附属医院眼科;063000 河北唐山，河北联合大学附属医院眼科;100000 北京，石景山区医院眼科;063000 河北唐山，河北联合大学附属医院眼科;063000 河北唐山，唐山市第九医院门诊部;063000河北唐山，河北联合大学附属医院眼科;063000 河北唐山，河北联合大学附属医院眼科【正文语种】中文【中图分类】R322.91【相关文献】1.神经生长因子联合银杏叶提取物对兔急性青光眼视网膜缺血再灌注损伤的保护作用 [J], 李岳美;李庆和;郑新华2.rh-bFGF 对兔视网膜缺血再灌注损伤的保护作用及其机制 [J], 邵宏超;葛嫣然;刘岩;苏杰;田华3.rh-bFGF对兔视网膜缺血再灌注损伤的保护作用及机制 [J], 邵宏超;葛嫣然;刘岩;苏杰;田华4.热休克蛋白72在兔视网膜缺血再灌注损伤中的表达及重组人碱性成纤维细胞生长因子对其表达的影响 [J], 邵宏超;葛嫣然;刘岩;苏杰;杨馥宇5.活血通络利水方对兔视网膜缺血/再灌注损伤的保护作用 [J], 杨赞章; 张越; 张铭连; 孟兢晶; 李明然因版权原因，仅展示原文概要，查看原文内容请购买。

腺病毒介导的碱性成纤维细胞生长因子对大鼠脊髓损伤腹后外侧索少突胶质细胞的影响杨彦玲【摘要】Objective To study the effect of adenovirus-mediated basic fibroblast growth factor ( FGF-2) gene transfer in vivo on oligodendrocyte cell numbers throughout ventrolateral white matter following spinal cord injury in rats. Methods Thirty-two adult female Sprague Dawley rats were injured with the Infinite Horizon Impactor, and then were randomly assigned to four groups; FGF-2-Adts high-titer group (1. 27 × 10 pfu/rat) , FGF-2 -Adts intermediate-titre group (6. 37 × 106 pfu/rat) , FGF-2-Adts low-titer group (3. 18 × 106 pfu/rat) , and green fluorescent protein (GFP) -Adts group (5. 9 × 107 pfu/rat). The transgenic expression in vivo was detected with fluorescence microscopy. The locomotor function of the hindlimbs of rats was evaluated using Riv-lin plate. Slides mounted with tissue sections were processed for immunohistochemical detection and quantification of oligodendrocytes (CC1 + ) in the ventral lateral funiculi (VLF) of injured spinal cords. Results One week after spinal cord injury, GFP showed that many cells had expressed objective gene in vivo and the angles of the occlusal plane of rats in FGF-2 groups were significantly higher than in GFP-Adts group. Also, there was a significant difference among the FGF-2-Adts treatment groups for the volume of gray matter sparing. However, there were no significant differences for total white matter sparing. Stereological quantification of total CC1 * cell numbers inthe spared VLF showed a significant reduction in numbers with GFP controls compared to all other groups 4 weeks after injury. In contrast, the FGF-2 Adts intermediate-titer group had significantly more CC1 + cells when compared to both the FGF-2-Adts high- and low-titer groups. Conclusion Adenovirus-mediated FGF-2 gene transfer can promote the functional recovery of the injured spinal cord by enhancing the proliferation and/or differentiation of oligodendrocytes.%目的观察腺病毒( Adts)介导碱性成纤维细胞生长因子(FGF-2)对大鼠脊髓损伤腹后外侧索少突胶质细胞的影响.方法 32只SD大鼠在T10水平制备脊髓损伤模型,随机分为体内转基因治疗组和损伤对照组,用携带FGF-2和绿色荧光蛋白(GFP)的Adts行直接体内转基因治疗.荧光显微镜观察体内转基因表达,斜板实验检测大鼠运动功能恢复情况,免疫组织化学染色观察腹后外侧索少突胶质细胞(CC1+)数量的变化.结果荧光显微镜观察发现,Adts载体直接引入脊髓后可有效感染脊髓组织并表达报告基因.斜板实验检测结果显示,体内转基因治疗组大鼠的倾斜平面角度明显高于对照组(P＜0.05或P＜0.01).免疫组织化学结果证实,腹后外侧索CCI+数量较对照组明显增加(P＜0.05).结论 Adts介导的FGF-2可促进少突胶质细胞的存活和增殖.【期刊名称】《中国医学科学院学报》【年(卷),期】2012(034)004【总页数】5页(P348-352)【关键词】脊髓损伤;碱性成纤维细胞生长因子;少突胶质细胞;腺病毒载体;基因治疗【作者】杨彦玲【作者单位】延安大学医学院生理教研室,陕西延安716000【正文语种】中文【中图分类】R744脊髓损伤是中枢神经系统的严重创伤，因其高致残率和高病死率一直成为医学研究的热门课题。

中国组织工程研究与临床康复 第15卷 第27期 2011–07–02出版Journal of Clinical Rehabilitative Tissue Engineering Research July 2, 2011 Vol.15, No.27P.O. Box 1200, Shenyang 110004 50481Department of Joint Surgery, 2Department of Rehabilitation, 3Central Laboratory, Affiliated Hospital, Qingdao University Medical College, Qingdao 266000, Shandong Province, ChinaSui Lai-jian ★,Master, Department of Joint Surgery, Affiliated Hospital, Qingdao Univ ersity Medical College, Qingdao 266000, Shandong Province, ChinaCorrespondence to: Wang Ying-zhen, Doctoral supervisor, Department of Joint Surgery, Affiliated Hospital, Qingdao University Medical College, Qingdao 266000, Shandong Province, China wangy ingzhenqd@ Supported by: the Science andTechnology Tackle Key Plan of Qingdao City, No.06-2-2-6- nsh-2*Received: 2011-01-03 Accepted: 2011-02-24转化生长因子ß3基因转染兔骨髓间充质干细胞诱导其向软骨表型的分化*★隋来健1，吕成昱1，张海宁1，徐 浩1，高正玉2，隋爱华3，王英振1Differentiation of rabbit mesenchymal stem cells into cartilage phenotype induced by transforming growth factor beta 3Sui Lai-jian 1, Lü Cheng-yu 1, Zhang Hai-ning 1, Xu Hao 1, Gao Zheng-yu 2, Sui Ai-hua 3, Wang Ying-zhen 1AbstractBACKGROUND: Endogenous induction of cartilage differentiation is to use a certain carrier to integrate the target genes into stem cells to induce self-differentiation by self-induced secretion of cytokines.OBJECTIVE: To observe the ability of rabbit bone marrow mesenchymal stem cells (BMSCs) transfected with transforming growth factor beta 3 (TGF-ß3) to differentiate into cartilage phenotype through adeno-associated virus vector.METHODS: The third passage BMSCs in logarithmic growth phase was transfected by rAAV2-TGF-ß3. Total protein was extracted from cells at 3, 6, 9, 12 days after transfection, and TGF-ß3 was detected through ELISA. After 2 weeks of culturing, mRNA expressions of type Ⅱ collagen were determined by RT -PCR and the collagen Ⅱ protein was detected by western blot. The expression of proteoglycan was shown by toluidine blue staining.RESULTS AND CONCLUSION: After transfection, BMSCs can express TGF-ß3 stably, and make a better cartilagetransformation compared to negative control group. TGF-ß3 can be transfected into BMSCs through adeno-associated virus vectors and induce cell differentiation into cartilage phenotype.Sui LJ, Lü CY, Zhang HN, Xu H, Gao ZY, Sui AH, Wang YZ. Differentiation of rabbit mesenchymal stem cells into cartilage phenotype induced by transforming growth factor beta 3.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2011;15(27): 5048-5052. [ ]摘要背景：内源性诱导软骨分为就是通过一定的载体将目的基因整合入干细胞内，使其自行分泌诱导因子诱导自身进行分化。

碱性成纤维细胞生长因子重组腺病毒转染兔骨髓间充质干细胞的表型变化蔡弢艺;陈雄生;贾连顺;孙延卿;林斌;陈长青【期刊名称】《中国组织工程研究》【年(卷),期】2014(000)023【摘要】BACKGROUND:Exogenous basic fibroblast growth factor (bFGF) plays an important role in the ligament tissue healing process, and the use of transgenic methods to transfect exogenous genes into cells can promote the secretion of bFGF. OBJECTIVE:To observe phenotypic changes and the bFGF protein expression after bFGF recombinant adenovirus was used to transfect rabbit bone marrow mesenchymal stem cells (BMSCs). METHODS:Passage 2 BMSCs were divided into three groups:Ad.bFGF-eGFP group, Ad.eGFP group and control group. Under a phase contrast microscope we observed the changes in cellmorphology. The expression of bFGF protein in BMSCs was determined by enzyme-linked immunosorbent assay (ELISA). The proliferative curve was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). RESULTS AND CONCLUSION:The transfected cells showed a uniform phenotype of fibroblasts. MTT colorimetric assay revealed that more proliferative activity of transfected BMSCs was shown in the&nbsp;Ad.bFGF-eGFP group than in the Ad.eGFP group and control group. ELISA results showed that expression of bFGF protein was higher in the Ad.bFGF-eGFP group than inthe Ad.eGFP group and control group (P<0.05). BFGF recombinant adenovirus can induce the differentiation of BMSCs into fibroblasts, increase proliferative ability and promote the expression of bFGF protein.%背景：外源性碱性成纤维细胞生长因子在韧带组织的愈合过程中发挥着重要作用，采用转基因方法将外源性基因转入细胞内能促进碱性成纤维细胞生长因子分泌。

碱性成纤维生长因子和人重组骨形态发生蛋白2诱导分化兔骨髓间充质干细胞胡运生;范清宇;马保安;张殿忠;刘云燕【期刊名称】《中国组织工程研究》【年(卷),期】2006(010)001【摘要】背景:骨髓间充质干细胞依赖特定的生长和分化因子能分化为骨、软骨、脂肪等不同组织.人重组骨形态发生蛋白2有明显的骨诱导作用,能诱导未分化的骨髓间充质干细胞不可逆地形成骨与软骨,并最终导致新骨生成;碱性成纤维生长因子亦参与调控骨髓间充质干细胞的软骨与骨的分化.目的:观察碱性成纤维生长因子和人重组骨形态发生蛋白2对兔骨髓间充质干细胞增殖和分化的影响,以便寻求合适的骨形成诱导方法以代替常规的成骨培养体系.设计:随机对照实验.单位:解放军第四军医大学唐都医院骨肿瘤研究所.材料:培养的兔骨髓间充质干细胞.方法:实验于2004-06/12在第四军医大学唐都医院骨肿瘤研究所暨全军骨科中心完成. ①体外培养的兔骨髓间充质干细胞分别经3种不同的生长因子(100μg/L人重组骨形态发生蛋白2,100μg/L碱性成纤维生长因子,100μg/L人重组骨形态发生蛋白2加100μg/L碱性成纤维生长因子)和成骨培养体系处理. ②其增殖和分化通过四甲基偶氮唑盐比色法,碱性磷酸酶、骨钙素检测和钙结节Von Kossa染色观察.主要观察指标:通过检测增殖率和碱性磷酸酶活性比较不同生长因子对兔骨髓间充质干细胞增殖和分化的影响.结果: ①人重组骨形态发生蛋白2能促进兔骨髓间充质干细胞的增殖和分化,特别是其分化. ②碱性成纤维生长因子能刺激兔骨髓间充质干细胞的增殖,与对照组相比,细胞增值率提高近100%;虽然对兔骨髓问充质干细胞分化无明显影响,但其能促进人重组骨形态发生蛋白2对兔骨髓间充质干细胞分化.结论:碱性成纤维生长因子和人重组骨形态发生蛋白2是兔骨髓间充质干细胞有效的分化因子,两者在体外都能促进兔骨髓间充质干细胞的增殖和分化.碱性成纤维生长因子和人重组骨形态发生蛋白2能协同作用加速骨诱导及骨形成,可以用作分化骨组织工程中的种子细胞.%BACKGROUND: Mesenchymal stem cells (MSCs) are capable of differentiating into a variety of lineages, including bone, cartilage, or fat, depending on the inducing stimuli, specific growth and differentiation factors.Recombinant human bone morphogenetic protein 2 (rhBMP-2) produced by gene engineering has an obvious osteoinductive activity and can induce undifferentiated mesenchymal cells into cartilage and bone irreversibly, resulting in new bone formation. Basic fibroblast growth factor (bFGF) modulates chondrogenic and osteogenic differentiation of MSCs.OBJECTIVE: To investigate the effect of bFGF and rhBMP-2 on the differentiation and proliferation of cultured rabbit mesenchymal stem cell in order to find out an optimal way of osteogenesis instead of conventional osteogenic supplements (OS).DESIGN: A randomized and controlled trial.SETTING: General Institute of Orthopaedic Oncology, Tangdu Hospital,Fourth Military Medical University of ChinesePLA.MATERIALS: The subjects were rabbit mesenchymal stem cells cultured by the author.METHODS: This experiment was conducted at the Center of Orthopaedic Surgery, General Institute of Orthopaedic Oncology, Tangdu Hospital,Fourth Military Medical University of Chinese PLA from June 2004 to December 2004. ①Rabbit MSCs cultured in vitro were treated with different growth factor (100 μg/L rhBMP-2, 100 μg/L bFGF, 10 μg/L rhBMP-2and 100 μg/L bFGF and OS; ②The proliferation and differentiationof MSCs were observed through activity of MTT, expression of alkaline phosphatase(ALP), and von Kossa staining.MAIN OUTCOME MEASURES: the rate of proliferation and the activity of ALP.RESULTS: ①rhBMP-2 could promote the proliferation and differentiation of MSCs, especially the cell differentiation; ②bFGF could stimulate the proliferation , the cellular proliferation rate increased 100% as compared with control group, and has no effect on differentiation of MSCs , but it could enhance effect on the cell proliferation of rhBMP-2.CONCLUSION: bFGF and rhBMP-2 are effective induction factors for MSCs. Both of them can stimulate the proliferation and differentiation of MSCs in vitro. bFGF and rhBMP-2 exerted a synergetic action in speeding up the pace of osteoinduction and osteogenesis and can be used to differentiate seed cells for tissue engineering bone.【总页数】3页(P163-165)【作者】胡运生;范清宇;马保安;张殿忠;刘云燕【作者单位】解放军第四军医大学唐都医院骨肿瘤研究所暨全军骨科中心,陕西省西安市,710038;解放军第四军医大学唐都医院骨肿瘤研究所暨全军骨科中心,陕西省西安市,710038;解放军第四军医大学唐都医院骨肿瘤研究所暨全军骨科中心,陕西省西安市,710038;解放军第四军医大学唐都医院骨肿瘤研究所暨全军骨科中心,陕西省西安市,710038;解放军第四军医大学唐都医院骨肿瘤研究所暨全军骨科中心,陕西省西安市,710038【正文语种】中文【中图分类】R394.2【相关文献】1.人骨形态发生蛋白2基因重组腺病毒表达载体转染兔骨髓间充质干细胞 [J], 何春耒;姬广林;刘午阳;吴东宝;何澄;王茂源;叶勇军;莫建文;高辉2.人骨形态发生蛋白2基因重组腺病毒表达载体转染兔骨髓间充质干细胞 [J], 何春耒;姬广林;刘午阳;吴东宝;何澄;王茂源;叶勇军;莫建文;高辉3.重组人甲状旁腺素对人骨髓间充质干细胞表达骨形态发生蛋白2的影响 [J], 李丁;王俊芳;张成玉4.骨形态发生蛋白2及碱性成纤维生长因子2对大鼠骨髓间充质干细胞膜片增殖和成骨分化的影响 [J], 张文静; 王佳; 田梦婷; 韩祥祯; 周琦琪; 何惠宇5.骨形态发生蛋白2及碱性成纤维生长因子2对大鼠骨髓间充质干细胞膜片增殖和成骨分化的影响 [J], 张文静; 王佳; 田梦婷; 韩祥祯; 周琦琪; 何惠宇因版权原因，仅展示原文概要，查看原文内容请购买。

兔骨髓间充质干细胞的生物学特征及其对不同生长因子的反应【摘要】为了研究兔骨髓间充质干细胞(rBM-MSC)的生物学特征及其对不同生长因子的反应，使用贴壁培养法从兔骨髓单个核细胞中获得间充质干细胞，在光学显微镜和电子显微镜下观察其基本生长行为和生物学特征；使用流式细胞仪检测rBM-MSC的免疫学表型；RT-PCR法检测其胶原表达；诱导rBM-MSC多向分化并使用特异性染色和RT-PCR予以鉴定；最后使用MTT法检测IL-1、3、8和HGF对rBM-MSC生长增殖的影响。

结果显示：贴壁生长的rBM-MSC具有典型的成纤维样细胞形态，可传15代以上，第5代rBM-MSC高表达基质受体CD44，低表达造血细胞标记CD45；RT-PCR显示高表达I型胶原，弱表达II型胶原，不表达X型胶原；在不同的诱导条件下，rBM-MSC 可被诱导分化为成骨细胞、软骨细胞、脂肪细胞和神经元样细胞。

rBM-MSC对IL-3最为敏感，10 ng/ml的低浓度IL-3可显着促进细胞增殖达32％以上，而高浓度IL-3能显着抑制其生长。

结论：分离培养了rBM-MSC，其生物学特征与人和猕猴等BM-MSC相似的生物学特性，低浓度IL-3可有效促进其增殖。

【关键词】骨髓；间充质干细胞；生长因子；兔Biological Characteristics of Rabbit Bone Marrow Mesenchymal Stem Cells and Their Response to Different Growth FactorsAbstractThis study was aimed to analyze the biological characteristics of rabbit bone marrow mesenchymal stem cells (rBM-MSCs) and their response to different growth factors. RabbitBM-MSCs were separated from bone marrow mononuclear cells by using adherent cultivation. Biologicalcharacteristics were investigated by optical and electron microscopy. Immunophenotype of rBM-MSCs was measured by flow cytometry. Theexpression of collagen was detected by RT-PCR. Differentiation potential was identified by specific staining andRT-PCR. The response of rBM-MSCs to IL-1, 3, 8 and HGF with different concentrations were tested by MTT. The results showed that the rBM-MSCs gave rise to a population of adherent cells characterized by the presence of a predominant cell type with a typical fibroblast-like morphology and could be cultured for over 15 passages. CD44 was highly expressed on F5 rBM-MSCs (32%) and CD45 was lowly expressed (%). Type I collagen was highly expressed, while type II collagen was lowly expressed and type X collagen was not detected on rBM-MSCs using RT-PCR method. In various conditions inducting differentiation, rBM-MSCs could differentiate into the osteoblast, chondrocyte, adipocyte and neuron-like cells. The rBM-MSCs weresensitive to IL-3, even low concentration (10 ng/ml) of IL-3 could promote the proliferation of rBM-MSCs effectively (32％, ), whereas high concentration IL-3 inhibited it significantly. It is concluded that rabbit BM-MSCs were successfully isolated and culture-expanded. The biological characteristics of rabbit BM-MSCs are similar to those of human and rhesus BM-MSCs. IL-3 with low concentration can promote the proliferation of rBM-MSCs effectively, but high concentration of IL-3 can inhibit their proliferation.Key wordsbone marrow; mesenchymal stem cell; growth factor; rabbit骨髓间充质干细胞具有高度自我更新和多向分化的潜能，在不同的诱导条件下，可分化为多种组织细胞类型，如骨髓基质细胞、成骨细胞、成软骨细胞和脂肪细胞［1］。

碱性成纤维细胞生长因子对小鼠骨髓内皮细胞增殖的促进作用王峰;张煜;李娜娜【期刊名称】《新乡医学院学报》【年(卷),期】2006(023)005【摘要】目的以内皮细胞培养基(Endo-M)培养小鼠骨髓内皮细胞,观察碱性成纤维细胞生长因子(bFGF)对骨髓内皮细胞增殖的影响.方法用Endo-M培养小鼠骨髓内皮细胞,通过Wright's Giemsa染色计数内皮细胞集落,免疫荧光法观察内皮细胞表型;加入不同浓度的bFGF,通过集落形成率、MTT法和流式细胞仪检测内皮细胞生长周期,观察bFGF对内皮细胞体外扩增的影响.结果 Endo-M 诱导的骨髓细胞可以生成内皮细胞,vWF检测阳性.加入不同浓度的bFGF,集落计数及MTT法均证实了bFGF对内皮细胞的增殖具有明显的促进作用,比较实验组(bFGF组)和对照组细胞的生长曲线,两者有明显差异(P＜0.05).结论 bFGF对内皮细胞的增殖具有促进作用.【总页数】3页(P475-477)【作者】王峰;张煜;李娜娜【作者单位】新乡医学院生理学与神经生物学教研室,河南,新乡,453003;新乡医学院生物化学与分子生物学教研室,河南,新乡,453003;新乡医学院人体解剖学教研室,河南,新乡,453003【正文语种】中文【中图分类】Q462;R331.2【相关文献】1.促进小鼠骨髓间充质干细胞增殖和成骨分化的血管内皮生长因子 [J], 张连方;康辉;邓廉夫;杨惠林2.小鼠骨髓间充质干细胞对人舌鳞状细胞癌Cal27细胞的归巢作用及其对Cal27细胞增殖和迁移的促进作用 [J], 孟琳;王璐;布文奂;丁鑫鑫;高华丽;王梓霖;刘玉兰;李琛;孙宏晨3.体外负压培养对小鼠骨髓间充质干细胞增殖能力和血管内皮生长因子分泌水平的影响 [J], 杨雄峰; 姜慧娇; 王小义; 郭黎姣; 周青; 韩欢欢; 李林林; 廖振宇; 雷振; 陈雪玲; 张宏伟; 吴向未4.体外负压培养对小鼠骨髓间充质干细胞增殖能力和血管内皮生长因子分泌水平的影响 [J], 杨雄峰; 陈雪玲; 张宏伟; 吴向未; 姜慧娇; 王小义; 郭黎姣; 周青; 韩欢欢; 李林林; 廖振宇; 雷振5.骨髓内皮细胞无血清条件培养液对骨髓内皮细胞增殖的促进作用 [J], 周小莹;王绮如;黄艳红;程腊梅;谭孟群因版权原因，仅展示原文概要，查看原文内容请购买。

基因重组腺病毒载体与慢病毒载体转染兔骨髓间充质干细胞的对比李诗鹏;李强;石正松;蔡伟良;宁寅宽;陶旋【摘要】背景:腺病毒载体和慢病毒载体均是较好的组织工程基因载体,两者在介导骨形态发生蛋白2转染兔骨髓间充质干细胞中的差异尚待探究.目的:比较腺病毒和慢病毒载体介导EGFP/骨形态发生蛋白2转染体外培养兔骨髓间充质干细胞的转导效率、持续时间及外源基因表达差异.方法:将第5代兔骨髓间充质干细胞分3组培养,A组以腺病毒载体介导EGFP/骨形态发生蛋白2(Ad-EGFP/BMP-2)转染细胞,B组以慢病毒载体介导EGFP/骨形态发生蛋白2(Lenti-EGFP/BMP-2)转染细胞,C组为未进行转染.转染24 h、48 h、72 h、1周、3周,检测EGFP基因表达;转染72 h后,免疫组织化学观察细胞骨形态发生蛋白2的表达;转染后72 h、1周、3周,Western blot检测骨形态发生蛋白2蛋白表达.结果与结论:①转染24 h后,A、B组可见EGFP表达,A组明显强于B组;转染48 h后,A、B组荧光进一步增强;转染72 h后,A组荧光强度近高峰,B组荧光持续增强.转染1周后,A组荧光强度开始下降,B组荧光强度仍然增强.转染3周后,A组荧光强度明显下降甚至消失,B组荧光强度有所下降,但仍然保持一定的表达.C组在各时间点均无EGFP表达;②A、B组胞浆均呈骨形态发生蛋白2阳性表达,C组呈阴性表达;③转染72 h后,A组骨形态发生蛋白2蛋白表达量强于B组;转染1周后,A组表达下降,B组表达增强并强于A 组;转染3周后,A组表达微弱,B组持续表达且明显强于A组;④结果表明,相对腺病毒载体,慢病毒载体介导EGFP/骨形态发生蛋白2基因转染兔骨髓间充质干细胞在表达持续时间上具有一定的优势.%BACKGROUND: Both lentiviral vector and adenoviral vector are considered as good vectors for gene mediation, and their differences in transferring bone morphogenetic protein 2 (BMP-2)into rabbit bone marrow mesenchymal stem cells (BMSCs) are unclear. OBJECTIVE: To compare the duration, efficiency and the deviation of exogenous gene expression after rabbit BMSCs transfection using lentiviral vector and adenoviral vector which are used to mediate enhanced green fluorescent proteins (EGFP) and BMP-2. METHODS: Rabbit BMSCs at passage 5 were exposed to Ad-EGFP-BMP-2 (group A) or Lenti-EGFP-BMP-2 (group B) with multiplicity of infection of 100, as transfection groups. And in control group (group C), the same quality of culture medium was required equivalent to the groups A and B. The expression of EGFP was observed by inverted fluorescence microscope at various time intervals. And the expression of exogenous gene BMP 2 in cells was detected and analyzed by immunohistochemical staining at 72 hours after transfection as well as by western blot at 72 hours, 1, 3 weeks after transfection. RESULTS AND CONCLUSION: The intense green fluorescence emerged under the microscope at 24-48 hours after transfection in group A, which was stronger than group B, reached the peak at 72 hours, and then decreased at 1 week until disappearance at 3 weeks. No EGFP expression was detected in group C. High expression of BMP-2 was found in group A but was dramatically downregulated after 1 week. Group B showed the high expression of EGFP/BMP-2 persisted for a longer period after transfected that even lasted for 3 weeks. Overall, the lentiviral vector and adenoviral vector can efficiently transfect rabbit BMSCs and stably express the target gene of EGFP/BMP-2. Under the same MOI, compared to the adenoviral vector, transfection of lentiviral vector to rabbit BMSCs is moreeffectively and expression of EGFP/BMP-2 can be persistent in a longer term.【期刊名称】《中国组织工程研究》【年(卷),期】2017(021)009【总页数】6页(P1340-1345)【关键词】干细胞;骨髓干细胞;慢病毒载体;腺病毒载体;骨髓间充质干细胞;基因转染;骨形态发生蛋白类;国家自然科学基金【作者】李诗鹏;李强;石正松;蔡伟良;宁寅宽;陶旋【作者单位】桂林医学院附属医院急诊创伤外科,广西壮族自治区桂林市 541001;桂林医学院附属医院急诊创伤外科,广西壮族自治区桂林市 541001;桂林医学院附属医院急诊创伤外科,广西壮族自治区桂林市 541001;桂林医学院附属医院急诊创伤外科,广西壮族自治区桂林市 541001;桂林医学院附属医院急诊创伤外科,广西壮族自治区桂林市 541001;桂林医学院附属医院急诊创伤外科,广西壮族自治区桂林市 541001【正文语种】中文【中图分类】R394.20 引言 Introduction骨组织工程在治疗骨科疾病如骨缺损等方面展现了良好的前景[1]。