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In vitro differentiation of adipose-derived stem cells and bone marrow-derived stromal stem cells into neuronal-like cells 被引量:21
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作者 Jin Zhou Guoping Tian +9 位作者 Jing'e Wang Xuefeng Cong Xingkai Wu Siyang Zhang Li Li Bing Xu Feng Zhu Xuedan Luo Jian Han Fengjie Hu 《Neural Regeneration Research》 SCIE CAS CSCD 2011年第19期1467-1472,共6页
Adipose-derived stem cells and bone marrow-derived stromal stem cells were co-cultured with untreated or Aβ1-40-treated PC12 cells, or grown in supernatant derived from untreated or Aβ1-40-treated PC12 cells. Analys... Adipose-derived stem cells and bone marrow-derived stromal stem cells were co-cultured with untreated or Aβ1-40-treated PC12 cells, or grown in supernatant derived from untreated or Aβ1-40-treated PC12 cells. Analysis by western blot and quantitative real-time PCR showed that protein levels of Nanog, Oct4, and Sox2, and mRNA levels of miR/125a/3p were decreased, while expression of insulin-like growth factor-2 and neuron specific enolase was increased. In comparison the generation of neuron specific enolase-positive cells was most successful when adipose-derived stem cells were co-cultured with Aβ1-40-treated PC12 cells. Our results demonstrate that adipose-derived stem cells and bone marrow-derived stromal stem cells exhibit trends of neuronal-like cell differentiation after co-culture with Aβ1-40-treated PC12 cells. This process may relate to a downregulation of miR-125a-3p mRNA expression and increased levels of insulin-like growth factor-2 expression. 展开更多
关键词 adipose-derived stem cells bone marrow-derived stromal stem cells DIFFERENTIATION NEURON miR-125a-3p neural regeneration
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Effects of corneal stromal cell-and bone marrow-derived endothelial progenitor cell-conditioned media on the proliferation of corneal endothelial cells 被引量:1
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作者 Meng-Yu Zhu Qin-Ke Yao +6 位作者 Jun-Zhao Chen Chun-Yi Shao Chen-Xi Yan Ni Ni Xian-Qun Fan Ping Gu Yao Fu 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2016年第3期332-339,共8页
AIM: To explore the effects of conditioned media on the proliferation of corneal endothelial cells (CECa) and to compare the efficiency of different conditioned media (CM). METHODS: Rat CECs, corneal stromal cel... AIM: To explore the effects of conditioned media on the proliferation of corneal endothelial cells (CECa) and to compare the efficiency of different conditioned media (CM). METHODS: Rat CECs, corneal stromal cells (CSCs), bone marrow -derived endothelial progenitor cells (BEPCs), and bone marrow-derived mesenchymal stem cells (BMSCs) were isolated and cultured in vitra CM was collected from CSCs, BEPCs, and BMSCSo CECs were cultivated in different culture media. Cell morphology was recorded, and gene and protein expression were analyzed.~ RESULTS: After grown in CM for 5d, CECs in each experimental group remained polygonal, in a cobblestone- like monolayer arrangement. Immunocytofluorescence revealed positive expression of Na+/K+-ATP, aquaporin 1 (AQP1), and zonula occludens 1 (ZO-1). Based on quantitative polymerase chain reaction (qPCR) analysis, Na +/K +-ATP expression in CSC-CM was notably upregulated by 1.3-fold (+0.036) (P〈0.05, n=3). The expression levels of ZO-1, neuron specific enolase (NSE), Vimentin, paired homebox 6 (PAX6), and procollagen type VII (COL8A1) were notably upregulated in each experimental group. Each CM had a positive effect on CEC proliferation, and CSC-CM had the strongest effect on proliferation.~ CONCLUSION: CSC-CM, BEPC-CM, and BMSC-CM not only stimulated the proliferation of CECs, but also maintained the characteristic differentiated phenotypes necessary for endothelial functions. CSC-CM had the most notable effect on CEC proliferation. KEYWORDS: conditioned medium; corneal endothelial cell; corneal stromal cell; bone marrow-derived endothelial progenitor cell; proliferation 展开更多
关键词 conditioned medium corneal endothelialcell corneal stromal cell bone marrow-derived endothelialprogenitor cell PROLIFERATION
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Mesenchymal Stromal Cells Derived from Human Embryonic Stem Cells, Fetal Limb and Bone Marrow Share a Common Phenotype but Are Transcriptionally and Biologically Different 被引量:2
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作者 Candida Vaz Betty Tan Bee Tee +2 位作者 Delicia Yong Qian Yi Lee Vivek Tanavde 《Stem Cell Discovery》 2017年第1期1-26,共26页
Mesenchymal stromal cells (MSCs) can be obtained from several sources and the significant differences in their properties make it crucial to investigate the differentiation potential of MSCs from different sources to ... Mesenchymal stromal cells (MSCs) can be obtained from several sources and the significant differences in their properties make it crucial to investigate the differentiation potential of MSCs from different sources to determine the optimal source of MSCs. We investigated if this biological heterogeneity in MSCs from different sources results in different mechanisms for their differentiation. In this study, we compared the gene expression patterns of phenotypically defined MSCs derived from three ontogenically different sources: Embryonic stem cells (hES-MSCs), Fetal limb (Flb-MSCs) and Bone Marrow (BM-MSCs). Differentially expressed genes between differentiated cells and undifferentiated controls were compared across the three MSC sources. We found minimal overlap (5% - 16%) in differentially expressed gene sets among the three sources. Flb-MSCs were similar to BM-MSCs based on differential gene expression patterns. Pathway analysis of the differentially expressed genes using Ingenuity Pathway Analysis (IPA) revealed a large variation in the canonical pathways leading to MSC differentiation. The similar canonical pathways among the three sources were lineage specific. The Flb-MSCs showed maximum overlap of canonical pathways with the BM-MSCs, indicating that the Flb-MSCs are an intermediate source between the less specialised hES-MSC source and the more specialised BM-MSC source. The source specific pathways prove that MSCs from the three ontogenically different sources use different biological pathways to obtain similar differentiation outcomes. Thus our study advocates the understanding of biological pathways to obtain optimal sources of MSCs for various clinical applications. 展开更多
关键词 Mesenchymal stromal cells (MSCs) human Embryonic Stem cells DERIVED MSCS (hES-MSCs) FETAL LIMB DERIVED MSCS (Flb-MSCs) bone Marrow DERIVED MSCS (BM-MSCs) Ontogenically DIFFERENT Sources Source Specific Canonical Pathways
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Human bone marrow stromal cells in cooperation with exogenous cytokines support in vitro expansion of cord blood CD34^+ cells
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《中国输血杂志》 CAS CSCD 2001年第S1期411-,共1页
关键词 bone human bone marrow stromal cells in cooperation with exogenous cytokines support in vitro expansion of cord blood CD34 cells CD
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Construction of Sox9 Gene Eukaryotic Expression Vector and Its Inductive Effects on Directed Differentiation of Bone Marrow Stromal Cells into Precartilaginous Stem Cells in Rats 被引量:1
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作者 胡伟华 郭风劲 +3 位作者 李锋 黄晖 张伟凯 陈安民 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2009年第3期291-295,共5页
Sox9 gene was cloned from immortalized precartilaginous stem cells and its eukaryotic expression vector constructed in order to explore the possibility of bone marrow-derived stromal cells differentiation into precart... Sox9 gene was cloned from immortalized precartilaginous stem cells and its eukaryotic expression vector constructed in order to explore the possibility of bone marrow-derived stromal cells differentiation into precartilaginous stem cells induced by Sox9. A full-length fragment of Sox9 was obtained by RT-PCR, inserted into pGEM-T Easy clone vector, and ligated with pEGFP-IRES2 expression vector by double digestion after sequencing. The compound plasmid was transfected into born marrow-derived stromal cells by Lipofectamine 2000, and the transfection efficacy and the expression of Sox9 and FGFR-3 were observed. Flow cytometry was used to identify the cell phenotype, and MTT was employed to assay proliferative viability of cells. Sequencing, restrictive endonuclease identification and RT-PCR confirmed that the expansion of Sox9 and construction of Sox9 expression vector were successful. After transfection of the recombinant vector into bone marrow-derived stromal cells, the expression of Sox9 and FGFR-3 was detected, and proliferative viability was not different from that of precartilaginous stem cells. It was concluded that Sox9 gene eukaryotic expression vector was successfully constructed, and the transfected bone marrow-derived stromal cells differentiated into the precartilaginous stem cells. 展开更多
关键词 precartilaginous stem cells SOX9 bone marrow-derived stromal cells TRANSFECTION
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Mesenchymal stem cells over-expressing hepatocyte growth factor improve small-for-size liver grafts regeneration 被引量:47
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作者 Yu, Y. Yao, A. H. Chen, N. Pu, L. Y. Fan, Y. Lv, L. Sun, B. C. Li, G. Q. Wang, X. H. 《南京医科大学学报(自然科学版)》 CAS CSCD 北大核心 2007年第10期1133-1133,共1页
关键词 间叶细胞 干细胞 肝细胞生长因子 肝移植 肝再生
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Rapid Communication:Preliminary study on the freeze-drying of human bone marrow-derived mesenchymal stem cells 被引量:1
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作者 Shao-zhi ZHANG Huan QIAN +6 位作者 Zhen WANG Ju-li FAN Qian ZHOU Guang-ming CHEN Rui LI Shan FU Jie SUN 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2010年第11期889-894,共6页
Long-term preservation and easy transportation of human bone marrow-derived mesenchymal stem cells(hBM-MSCs) will facilitate their application in medical treatment and bioengineering.A pilot study on the freeze-drying... Long-term preservation and easy transportation of human bone marrow-derived mesenchymal stem cells(hBM-MSCs) will facilitate their application in medical treatment and bioengineering.A pilot study on the freeze-drying of hBM-MSCs was carried out.hBM-MSCs were loaded with trehalose.The glass transition temperature of the freeze-drying suspension was measured to provide information for the cooling and primary drying experiment.After freeze-drying,various rehydration processes were tested.The highest recovery rate of hBM-MSCs was(69.33± 13.08) %.Possible methods to improve freeze-drying outcomes are discussed.In conclusion,the present study has laid a foundation for the freeze-drying hBM-MSCs. 展开更多
关键词 human bone marrow-derived mesenchymal stem cells (hBM-MSCs) FREEZE-DRYING TREHALOSE REHYDRATION
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Therapeutic Approach for Hair Growth and Regeneration Using Bioactive Formulation Containing Mesenchymal Stromal Cell-Derived Conditioned Medium
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作者 Pawan Kumar Gupta Samatha Bhat +2 位作者 Suresh Kannan Raviraja N. Seetharam Udaykumar Kolkundkar 《Journal of Cosmetics, Dermatological Sciences and Applications》 2023年第3期182-208,共27页
Background and Aims: Androgenetic alopecia (AGA) is a common form of hair loss in both men and women. Despite its high prevalence and associated patient morbidity, the approved therapeutic options are limited to finas... Background and Aims: Androgenetic alopecia (AGA) is a common form of hair loss in both men and women. Despite its high prevalence and associated patient morbidity, the approved therapeutic options are limited to finasteride and minoxidil. The present study is aimed at assessing the efficacy of hair serum formulation, Trichosera<sup>®</sup>containing Bone marrow-derived mesenchymal stromal cells conditioned media as an active ingredient, for hair fall control and hair regrowth in healthy Indian human volunteers. Methods: The product was made using a 20% concentration of 10X Conditioned Media along with excipients. The final product was tested for physicochemical parameters, biomarkers, total protein content and microbial limits as per our in-house specifications. Results: The primary irritation patch test showed that the product is non-irritant and dermatologically safe. A clinical study on 40 subjects was conducted to evaluate the effectiveness of the bioactive formulation in hair fall control and hair regrowth in healthy volunteers. Phototrichogram measurement showed hair density and hair growth rate increased significantly by 11.54% and 18.66% at week 24. Hair tensile strength also increased significantly by 41.10% at 12 weeks follow-up. Hair pull test, to see a reduction in pulled hair and comb’s test to show a decrease in hair fall significantly improved from week 4 onwards. There were no significant adverse events in response to the product application. Conclusion: It is concluded that the hair serum product is completely safe on direct application to the scalp and showed significant improvement in the hair growth rate, hair density, scalp condition and reduction in hair fall. . 展开更多
关键词 Trichosera® bone Marrow Derived Mesenchymal stromal cells Conditioned Media Hair Fall Hair Regrowth human Volunteer Study
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Sustained adenosine release:Revealing its impact on osteogenic signalling pathways of human mesenchymal stromal cells
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作者 Hadi Hajiali Jane McLaren +5 位作者 Cristina Gonzalez-García Salah Abdelrazig Dong-Hyun Kim Matthew J.Dalby Manuel Salmerón-Sánchez Felicity R.A.J.Rose 《Engineered Regeneration》 EI 2024年第2期255-268,共14页
Non-healing fractures,a global health concern arising from trauma,osteoporosis,and tumours,can lead to severe disabilities.Adenosine,integral to cellular energy metabolism,gains prominence in bone regeneration via ade... Non-healing fractures,a global health concern arising from trauma,osteoporosis,and tumours,can lead to severe disabilities.Adenosine,integral to cellular energy metabolism,gains prominence in bone regeneration via adeno-sine A2 B receptor activation.This study introduces a controlled-release system for localized adenosine delivery,fostering human mesenchymal stromal cell(hMSC)differentiation into functional bone cells.The study investi-gates how the ratio of lactic acid to glycolic acid in microparticles can influence adenosine release and explores the downstream effects on gene expression and metabolic profiles of osteogenic differentiation in hMSCs cultured in growth and osteoinductive media.Insights into adenosine-modulated signalling pathways during MSC differenti-ation,with osteogenic factors,provide a comprehensive understanding of the pathways involved.Analysing gene expression and metabolic profiles unravels adenosine’s regulatory mechanisms in MSC differentiation.Sustained adenosine release from microparticles induces mineralization,synergizing with osteogenic media supplements,showcasing the potential of adenosine for treating critical bone defects and metabolic disorders.This study high-lights the efficacy of a polymeric microparticle-based delivery system,offering novel strategies for bone repair.Unveiling adenosine’s roles and associated signalling pathways advances our comprehension of molecular mech-anisms steering bone regeneration,propelling innovative biomaterial,combined with metabolites,approaches for clinical use. 展开更多
关键词 Adenosine release Osteogenic signalling pathways human mesenchymal stromal cells bone regeneration
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Umbilical cord-derived mesenchymal stromal cells:Promising therapy for heart failure
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作者 Ya-Lun Li En-Guo Chen Bing-Bing Ren 《World Journal of Cardiology》 2025年第1期15-21,共7页
Heart failure(HF)is a complex syndrome characterized by the reduced capacity of the heart to adequately fill or eject blood.Currently,HF remains a leading cause of morbidity and mortality worldwide,imposing a substant... Heart failure(HF)is a complex syndrome characterized by the reduced capacity of the heart to adequately fill or eject blood.Currently,HF remains a leading cause of morbidity and mortality worldwide,imposing a substantial burden on global healthcare systems.Recent advancements have highlighted the therapeutic potential of mesenchymal stromal cells(MSCs)in managing HF.Notably,umbilical cord-derived MSCs(UC-MSCs)have demonstrated superior clinical potential compared to traditional bone marrow-derived MSCs;this is evident in their non-invasive collection process,higher proliferation efficacy,and lower immunogenicity and tumorigenicity,as substantiated by preclinical studies.Although the feasibility and safety of UC-MSCs have been tested in animal models,the application of UC-MSCs in HF treatment remains challenged by issues such as inaccurate targeted migration and low survival rates of UC-MSCs.Therefore,further research and clinical trials are imperative to advance the clinical application of UC-MSCs. 展开更多
关键词 Umbilical cord-derived mesenchymal stromal cells bone marrow-derived mesenchymal stromal cells cell therapy Heart failure Cardiovascular diseases
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基于UHPLC-Q-TOF/MS技术分析肿节风总黄酮提取物促进巨核细胞分化的效应成分
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作者 张钟康 卢晓南 +4 位作者 卢震 胡佳 刘慧珍 卢婷 尚广彬 《中药新药与临床药理》 CAS CSCD 北大核心 2024年第1期56-64,共9页
目的筛选肿节风总黄酮提取物促进巨核细胞分化的效应成分。方法(1)以人巨核细胞白血病细胞(Dami)与人骨髓基质细胞(HS-5)共培养的方式建立巨核细胞分化障碍模型作为评价体系,实验分组:Dami组(Dami)、对照组(Dami+HS-5)、PMA组[Dami+HS-5... 目的筛选肿节风总黄酮提取物促进巨核细胞分化的效应成分。方法(1)以人巨核细胞白血病细胞(Dami)与人骨髓基质细胞(HS-5)共培养的方式建立巨核细胞分化障碍模型作为评价体系,实验分组:Dami组(Dami)、对照组(Dami+HS-5)、PMA组[Dami+HS-5+5 ng·mL-1佛波醇12-十四酸酯13-乙酸酯(PMA)]、模型组[Dami+HS-5+1%兔抗大鼠血小板血清(APS)+5 ng·mL-1PMA],培养48 h。采用流式细胞术检测巨核细胞分化成熟表面标记分子CD41a、CD61的表达情况。(2)将49只SD雄性大鼠随机分为空白血浆组、15 min组、30 min组、60 min组、90 min组、120 min组、240 min组,每组7只。各给药组大鼠灌胃肿节风总黄酮提取物1.26 g·kg^(-1),在6个设定时间点(15、30、60、90、120、240 min)采血制备肿节风总黄酮提取物经时含药血浆。(3)采用超高效液相色谱-四极杆串联飞行时间质谱法(UHPLC-Q-TOF/MS)对肿节风总黄酮提取物经时含药血浆进行分析,以峰面积构建肿节风总黄酮提取物经时含药血浆中的化学成分随时间变化量矩阵(X矩阵)。将所采集的6个不同时间点的肿节风总黄酮经时含药血浆对巨核细胞分化成熟障碍模型进行干预,采用流式细胞术检测细胞表面分子CD41a、CD61的表达水平,构建肿节风总黄酮提取物经时含药血浆效应矩阵(Y矩阵)。(4)将X矩阵和Y矩阵数据标准化处理后,采用偏最小二乘法(Partial least squares,PLS)计算分析量效关系,以变量重要性投影(Variable importance for projection,VIP)>1为阈值,筛选与细胞表面分子CD41a、CD61变化相关的效应成分,并进行化学成分鉴定,作为肿节风总黄酮提取物中促进巨核细胞分化的潜在效应成分,最后回归评价体系验证其药效。结果(1)与Dami组比较,对照组Dami细胞表面的CD41a表达水平明显升高(P<0.05)。与对照组比较,PMA组Dami细胞表面的CD41a、CD61表达水平均显著升高(P<0.01)。与PMA组比较,模型组Dami细胞表面的CD41a、CD61表达水平均显著降低(P<0.01)。(2)与空白血浆组比较,15、30、60、90、120、240 min各时间点Dami细胞表面分子CD41a、CD61表达水平均显著升高(P<0.01),且CD41a、CD61均在30 min组表达水平最高。在正、负离子模式下筛选出VIP值>1的潜在效应成分,并选取540.3638@12.25与559.2991@11.53两个成分进行药效学验证。559.2991@11.53被鉴定为胡萝卜苷(Daucosterol,Dau),540.3638@12.25被鉴定为迷迭香酸-4-O-β-D-葡萄糖(Rosmarinic acid 4-O-β-Dglucoside,Ros)。Ros、Dau分别干预巨核细胞分化成熟障碍模型后,与模型组比较,Ros及Dau低、中、高剂量组(40、60、80μg·mL-1)的Dami细胞表面的CD41a、CD61表达水平均显著升高(P<0.05,P<0.01)。结论Ros、Dau可能是肿节风总黄酮提取物促进巨核细胞分化的效应成分。 展开更多
关键词 肿节风总黄酮提取物 免疫性血小板减少症 巨核细胞分化障碍模型 人巨核白血病细胞 人骨髓基质细胞 超高效液相色谱-四极杆串联飞行时间质谱法 偏最小二乘法 胡萝卜苷 迷迭香酸-4-O-β-D-葡萄糖
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PPARγ基因沉默的人骨髓基质细胞对骨髓抑制小鼠造血功能的影响
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作者 王雪梅 黄纯兰 +5 位作者 周铁军 李玉娇 魏梦宇 陈燕 陈晓敏 王万玥 《华中科技大学学报(医学版)》 CAS CSCD 北大核心 2024年第1期6-12,共7页
目的 观察过氧化物酶体增殖激活受体γ(peroxisome proliferator activated receptor-gamma, PPARγ)基因沉默的人骨髓基质细胞HS-5对骨髓抑制小鼠造血功能的影响,并初步探讨其可能的作用机制。方法 用X射线进行全身照射构建骨髓抑制小... 目的 观察过氧化物酶体增殖激活受体γ(peroxisome proliferator activated receptor-gamma, PPARγ)基因沉默的人骨髓基质细胞HS-5对骨髓抑制小鼠造血功能的影响,并初步探讨其可能的作用机制。方法 用X射线进行全身照射构建骨髓抑制小鼠模型,造模后2 h,将小鼠随机分为3组,分别为实验组(尾静脉注射PPARγ RNAi干扰的HS-5细胞)、对照组(尾静脉注射未行PPARγ RNAi干扰的HS-5细胞)、空白组(尾静脉注射等量的生理盐水),每组5只。各组于放疗前、放疗后24 h、放疗后1周、放疗后2周进行外周血常规检测。对HS-5细胞在体外进行成骨、成脂诱导,分为实验组(PPARγ RNAi干扰的HS-5细胞)、对照组(未干扰PPARγ的HS-5细胞)、空白组(未行成骨/成脂诱导分化的HS-5细胞),观察成骨/成脂染色情况。采用CCK-8实验检测PPARγ基因沉默的HS-5细胞对小鼠骨髓造血干细胞(hemopoietic stem cell, HSC)增殖的影响,分为实验组(PPARγ RNAi干扰的HS-5细胞经成骨诱导分化3 d后,与小鼠HSC共培养)、阳性对照组(50μmol/L PPARγ抑制剂处理的HS-5细胞经成骨诱导分化3 d后,与小鼠HSC共培养)、阴性对照组(未干扰PPARγ的HS-5细胞经成骨诱导分化3 d后,与小鼠HSC共培养)、空白组(小鼠HSC单独培养,不与HS-5细胞共培养)。结果 放疗后,各组小鼠血常规指标均呈先降低后升高趋势,放疗后1周,三组小鼠血小板、白细胞水平差异显著,且实验组>对照组>空白组(均P<0.05);放疗后2周,三组小鼠脂肪空泡面积百分比差异显著,且实验组<对照组<空白组(均P<0.05),经Pearson相关分析显示,血常规各指标与血清PPARγ表达水平呈负相关(均P<0.05),与脂肪空泡面积百分比呈负相关(均P<0.05)。在体外成骨/成脂诱导分化后,实验组与对照组相比,橙红色的细胞比例明显降低,红色钙结节比例明显增高;成骨分化诱导3 d后,实验组、阳性对照组、阴性对照组人骨髓基质细胞均与小鼠HSC细胞进行共培养,空白组则单纯培养HSC细胞,结果显示共培养24、48、72 h后,实验组、阳性对照组小鼠HSC细胞增殖水平均高于阴性对照组和空白组(均P<0.05)。结论 PPARγ基因沉默的HS-5植入骨髓抑制小鼠后有助于小鼠造血功能增强。PPARγ基因被干扰沉默后,可增强HS-5细胞的成骨分化能力,减弱HS-5细胞的成脂分化能力,而成骨分化诱导的HS-5细胞能进一步增强小鼠HSC的增殖能力。 展开更多
关键词 过氧化物酶体增殖激活受体Γ 人骨髓基质细胞 骨髓抑制小鼠 造血功能 成骨/成脂分化
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人骨形态发生蛋白-2腺病毒表达载体转染人骨髓基质干细胞及对其增殖的影响 被引量:7
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作者 李建军 刘建国 +5 位作者 韩东 孙洪斌 卜丽莎 杨绍娟 张文岚 徐莘香 《吉林大学学报(医学版)》 CAS CSCD 北大核心 2003年第4期421-424,共4页
目的 :用人骨形态发生蛋白 - 2腺病毒表达载体 ( Ad- BMP- 2 )转染人骨髓基质干细胞( h BMSC) ,以探讨基因转染对 h BMSC增殖的影响。方法 :将 Ad- BMP- 2转染体外培养的成人骨髓基质干细胞 ,利用免疫组化、原位杂交染色方法检测细胞内... 目的 :用人骨形态发生蛋白 - 2腺病毒表达载体 ( Ad- BMP- 2 )转染人骨髓基质干细胞( h BMSC) ,以探讨基因转染对 h BMSC增殖的影响。方法 :将 Ad- BMP- 2转染体外培养的成人骨髓基质干细胞 ,利用免疫组化、原位杂交染色方法检测细胞内骨形态发生蛋白 - 2 ( BMP- 2 )的表达 ,蛋白印迹法检测转染后细胞培养液中 BMP- 2分泌蛋白表达。然后通过流式细胞仪分析其对细胞增殖的影响。结果 :转染后 ,h BMP- 2基因在 m RNA水平和蛋白水平均有表达。蛋白印迹法检测到培养液中有 BMP- 2蛋白阳性表达。转染 h BMP- 2基因后 ,细胞经流式细胞仪分析 ,S期细胞比例增多 ,说明细胞 DNA的合成增加。结论 :Ad- BMP- 2可高效转染 h BMSC。 展开更多
关键词 腺病毒 骨形态发生蛋白 骨髓基质干细胞 基因转染
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人胚神经干细胞定向诱导分化为多巴胺能神经元的实验研究 被引量:18
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作者 张力 江澄川 +3 位作者 徐斌 陈商群 冯林音 江澄川 《中国神经精神疾病杂志》 CAS CSCD 北大核心 2002年第6期410-413,共4页
目的 建立神经干细胞与骨髓基质细胞的共培养系统,根据该系统的条件性培养液诱导多巴胺能神经元的分化。方法 来源于胎脑海马、纹状体、额叶、中脑的神经干细胞与骨髓基质细胞建立起各自的共培养系统,并根据数种条件性培养液诱导神经... 目的 建立神经干细胞与骨髓基质细胞的共培养系统,根据该系统的条件性培养液诱导多巴胺能神经元的分化。方法 来源于胎脑海马、纹状体、额叶、中脑的神经干细胞与骨髓基质细胞建立起各自的共培养系统,并根据数种条件性培养液诱导神经干细胞的分化,以免疫细胞化学检测神经元的总体分化率及多巴胺能神经元的诱导率。结果 骨髓基质细胞及CO-BMSC能显著提高不同来源的神经干细胞的神经元分化率,同时只有中脑神经干细胞能被有效地进行多巴胺能神经元的诱导。结论 共培养系统诱发了神经干细胞与骨髓基质细胞的自/旁分泌作用,该作用可根据神经干细胞的区域特异性有效的定向诱导中脑神经干细胞的分化。 展开更多
关键词 人胚神经干细胞 定向诱导分化 实验研究 骨髓基质细胞 多巴胺能神经元
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人脐血基质细胞自发分泌多种造血生长因子的实验研究 被引量:8
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作者 刘耀 陈幸华 +1 位作者 张曦 彭贤贵 《第三军医大学学报》 CAS CSCD 北大核心 2005年第7期643-645,共3页
目的 探讨体外培养的人脐血基质细胞分泌血小板生成素(thrombopoietin ,TPO)、粒 巨噬细胞集落刺激因子(granulocyte macrophagecolonystimulatingfactor,GM CSF)、干细胞生长因子(stemcellfactor ,SCF)的能力。方法 留取不同培养时... 目的 探讨体外培养的人脐血基质细胞分泌血小板生成素(thrombopoietin ,TPO)、粒 巨噬细胞集落刺激因子(granulocyte macrophagecolonystimulatingfactor,GM CSF)、干细胞生长因子(stemcellfactor ,SCF)的能力。方法 留取不同培养时相点的培养液上清,ELISA法检测上清中TPO、GM CSF、SCF的浓度,并与骨髓基质细胞的培养上清液对照比较其特点。结果 人脐血基质细胞和骨髓基质细胞均可分泌TPO、GM CSF和SCF等造血生长因子,二者造血生长因子的分泌随着培养时间的延长而逐渐下降,其中人脐血基质细胞分泌TPO和SCF的高峰期出现在第7天,而GM CSF的分泌则无高峰期的出现;通过与骨髓基质细胞分泌三种造血生长因子量进行对比发现,人脐血基质细胞SCF和GM CSF的分泌量低于骨髓基质细胞,而TPO的分泌高于同期培养的骨髓基质细胞。结论 人脐血基质细胞与骨髓基质细胞一样具有分泌造血生长因子,支持调控造血的作用。 展开更多
关键词 人脐血基质细胞 造血生长因子 骨髓基质细胞
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茶黄素对人骨髓基质干细胞的成骨诱导作用 被引量:4
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作者 尚希福 路玉峰 +5 位作者 张文志 张梅 贺瑞 姚刚 胡飞 曾建学 《山东医药》 CAS 北大核心 2010年第19期7-9,共3页
目的探讨茶黄素对体外培养人骨髓基质干细胞(BMSCs)的成骨诱导作用及其能力。方法将第二代人BMSCs分为茶黄素组、对照组,茶黄素组加入浓度为10 mmol/L茶黄素,对照组不干预。对比观察两组干预后细胞形态,钙结节、碱性磷酸酶(ALP)、Ⅰ型... 目的探讨茶黄素对体外培养人骨髓基质干细胞(BMSCs)的成骨诱导作用及其能力。方法将第二代人BMSCs分为茶黄素组、对照组,茶黄素组加入浓度为10 mmol/L茶黄素,对照组不干预。对比观察两组干预后细胞形态,钙结节、碱性磷酸酶(ALP)、Ⅰ型胶原及骨钙素(OCN)免疫组化染色结果,并检测各组4、8、12、16 d各时点ALP、OCN活性。结果茶黄素组钙结节、ALP、Ⅰ型胶原及OCN免疫组化染色均呈阳性,对照组均为阴性;茶黄素组各时点ALP、OCN活性均高于对照组(P均<0.05)。结论茶黄素可促进人BMSCs向成骨细胞分化。 展开更多
关键词 人骨髓基质干细胞 成骨细胞 诱导 茶黄素 骨钙素
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多发性骨髓瘤骨髓基质细胞IL-6和TNF-α的表达及其临床意义 被引量:5
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作者 高力 高蕾 +1 位作者 张曦 陈幸华 《重庆医学》 CAS CSCD 北大核心 2009年第7期801-802,810,共3页
目的研究骨髓瘤骨髓基质细胞(BMSCs)分泌的白介素-6(IL-6)和肿瘤坏死因子α(TNF-α)在多发性骨髓瘤(MM)进展中的变化和意义以及人脐血源基质细胞(hUCBDSCs)分泌IL-6和TNF-α的水平。方法用ELISA法检测单独培养的hUCBDSCs和骨髓瘤BMSCs... 目的研究骨髓瘤骨髓基质细胞(BMSCs)分泌的白介素-6(IL-6)和肿瘤坏死因子α(TNF-α)在多发性骨髓瘤(MM)进展中的变化和意义以及人脐血源基质细胞(hUCBDSCs)分泌IL-6和TNF-α的水平。方法用ELISA法检测单独培养的hUCBDSCs和骨髓瘤BMSCs以及它们与KM3细胞共培养后IL-6、TNF-α在培养上清液中的浓度。结果(1)单独培养和共培养时骨髓瘤BMSCs分泌IL-6和TNF-α的水平高于hUCBDSCs;(2)共培养后骨髓瘤BMSCs和hUCBDSCs分泌IL-6和TNF-α的水平明显高于共培养前;(3)Ⅲ期患者BMSCs分泌IL-6的水平高于Ⅰ期和Ⅱ期患者,但是TNF-α水平无变化。结论IL-6与MM的进展有关;hUCBDSCs表达IL-6和TNF-α的水平较骨髓瘤BMSCs低。 展开更多
关键词 多发性骨髓瘤 人脐血源基质细胞 骨髓基质细胞 白介素-6 肿瘤坏死因子Α
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骨髓基质细胞转化成骨细胞修复骨缺损:成纤维细胞生长因子和骨形成蛋白-2的作用 被引量:7
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作者 张晓东 杨维东 +2 位作者 顾晓明 钱奇春 赵丽君 《中国组织工程研究与临床康复》 CAS CSCD 2001年第20期44-45,共2页
目的探讨成纤维细胞生长因子( bFGF)和重组人骨形成蛋白- 2( rhBMP- 2)单独和联合作用对骨髓基质细胞( BMSC)的增殖和分化的影响。方法利用四唑盐比色法( MTT),细胞总蛋白考马斯亮蓝测定法,碱性磷酸酶( ALP)试剂盒分别测... 目的探讨成纤维细胞生长因子( bFGF)和重组人骨形成蛋白- 2( rhBMP- 2)单独和联合作用对骨髓基质细胞( BMSC)的增殖和分化的影响。方法利用四唑盐比色法( MTT),细胞总蛋白考马斯亮蓝测定法,碱性磷酸酶( ALP)试剂盒分别测定不同浓度的 bFGF和 rhBMP单独和联合作用后 BMSC的增殖和分化情况。结果 bFGF单独作用促进 BMSC的增殖,但高浓度抑制 ALP表达, rhBMP对 BMSC的增殖和 ALP和总蛋白含量均有促进作用,而 bFGF和 rhBMP联合作用后 BMSC的增殖和 ALP活性较单独作用有更明显的升高。结论 bFGF和 rhBMP联合作用对于促进 BMSC的增殖和向成骨细胞分化有协同作用。 展开更多
关键词 骨髓基质细胞 碱性成纤维细胞生长因子 骨形成蛋白 增殖 分化
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rhBMP-2体外诱导骨质疏松大鼠BMSCs成骨及VEGF表达的研究 被引量:12
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作者 李军 王云 +2 位作者 鲍小明 卫鹏斌 张民 《中国骨伤》 CAS 2015年第5期446-449,共4页
目的 :观察骨形态发生蛋白-2对骨质疏松时骨髓基质干细胞(BMSCs)体外成骨及成骨因子VEGF表达的影响,为骨质疏松证的防治提供新的方法。方法:将20只6月龄,体重(300±20)g雌性SD大鼠双侧卵巢切除,术后3个月利用双能X线骨密度仪测量大... 目的 :观察骨形态发生蛋白-2对骨质疏松时骨髓基质干细胞(BMSCs)体外成骨及成骨因子VEGF表达的影响,为骨质疏松证的防治提供新的方法。方法:将20只6月龄,体重(300±20)g雌性SD大鼠双侧卵巢切除,术后3个月利用双能X线骨密度仪测量大鼠全身骨密度并与术前比较,确保造模成功,并运用全骨髓贴壁法培养骨质疏松大鼠BMSCs,倒置相差显微镜下观察BMSCs形态。随机把骨质疏松大鼠BMSCs第2代(p2)细胞分成实验组和对照组,分别加入完全培养基(含rh BMP-2)、成骨诱导液进行成骨诱导。2周后茜素红染色法检测各组细胞钙结节的形成,酶标仪测定碱性磷酸酶活性及RT-PCR法检测VEGF的表达量。结果:(1)大鼠全身骨密度:手术前后大鼠全身骨密度分别为(0.179±0.007),(0.158±0.006)g/cm2,差异有统计学意义(t=4.180,P<0.05)。(2)茜素红染色:BMSCs(P2)成骨诱导2周后实验组染色效果明显强与对照组。(3)碱性磷酸酶活性:BMSCs(P2)成骨诱导2周后碱性磷酸酶活性实验组明显高于对照组,分别为(15.62±1.27),(8.62±0.93)μg/prot,差异有统计学意义(t=7.709,P<0.01)。(4)BMSCs(P2)成骨诱导2周后VEGF表达:实验组明显高于对照组,分别为3.723±0.143,0.950±0.072,差异有统计学意义(t=29.462,P<0.01)。结论 :rh BMP-2能提高去卵巢骨质疏松大鼠BMSCs的体外成骨能力,可促进成骨因子VEGF的表达,调控VEGF的表达可能是骨形态发生蛋白-2参与骨代谢的机制之一。 展开更多
关键词 重组人骨形态发生蛋白-2 骨质疏松 血管内皮因子 骨髓基质干细胞
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人骨髓基质细胞接种珊瑚构建组织工程骨 被引量:3
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作者 钱奇春 刘彦普 +3 位作者 杨维东 陈富林 赵黎 张晓东 《中国矫形外科杂志》 CAS CSCD 2002年第4期360-361,共2页
目的 :观察体外培养的人骨髓基质细胞与珊瑚复合物植入体内后的成骨能力。方法 :穿刺抽吸人髂骨区骨髓基质细胞 ,体外培养扩增、诱导 ,将其与珊瑚复合后植入裸鼠体内 ,以单纯珊瑚作为对照。术后 4、8周取材 ,通过大体、组织学、扫描电... 目的 :观察体外培养的人骨髓基质细胞与珊瑚复合物植入体内后的成骨能力。方法 :穿刺抽吸人髂骨区骨髓基质细胞 ,体外培养扩增、诱导 ,将其与珊瑚复合后植入裸鼠体内 ,以单纯珊瑚作为对照。术后 4、8周取材 ,通过大体、组织学、扫描电镜观察植入物体内成骨情况。结果 :术后 4周 ,复合物中有少量新骨形成 ;术后 8周 ,复合物中出现大量成熟骨组织。而对照组无骨组织形成。结论 :人骨髓基质细胞复合珊瑚在无免疫动物体内具有成骨能力 。 展开更多
关键词 人骨髓基质细胞 珊瑚 成骨 骨组织工程
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