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Expression of Caspase-3 in Cord Blood CD34^+ Cells during Culture in vitro
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作者 马艳萍 邹萍 +1 位作者 肖娟 黄士昂 《The Chinese-German Journal of Clinical Oncology》 CAS 2003年第3期166-168,192,共4页
Objective: To investigate the expression and significance of caspase-3 protein in CD34^+ cells from cord blood (CB) during culture in vitro with different growth factors. Methods: RT-PCR, Western blot and flow cytomet... Objective: To investigate the expression and significance of caspase-3 protein in CD34^+ cells from cord blood (CB) during culture in vitro with different growth factors. Methods: RT-PCR, Western blot and flow cytometry techniques were used to detect the expression of caspase-3 in CD34^+ CB cells during culture in vitro. Results: Caspase-3 mRNA was constitutively expressed at a low level in freshly isolated CD34^+ cells. The expression of caspase-3 mRNA and protein was upregulated when these cellswere first expanded in suspension culture with growth factors for 3 days. However, only the 32 kDa inactive caspase-3 proenzyme was detected in the freshly isolated CD34^+ cells as well as during the first 3 days expansion with cytokines. With longer culture time in vitro, especially in the presence of the combination of IL-3, IL-6 and GM-CSF, caspase-3 was activated and a cleavage product of 20 kDa became detectable.Conclusion: Caspase-3 is involved in apoptosis of primitive CB CD34^+ cells during expansion in vitro. 展开更多
关键词 CASPASE-3 ^cd34^+ cells cord blood APOPTOSIS
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THE EFFECT OF STEM CELL FACTOR, INTERLEUKIN-6 AND ERYTHROPOIETIN ON EXPANSION OF CD34^+ CELLS FROM HUMAN UMBILICAL CORD BLOOD
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作者 隋星卫 《中国实验血液学杂志》 CAS CSCD 1995年第4期390-394,共5页
CD34+ cells from human umbilical cord blood were purified by Dynal beads M-450 CD34 immunoselection system and cultured in the presence of various cytokines alone or in combination, including stem cell factor (SCF), i... CD34+ cells from human umbilical cord blood were purified by Dynal beads M-450 CD34 immunoselection system and cultured in the presence of various cytokines alone or in combination, including stem cell factor (SCF), interleukin-6 (IL-6) and erythropoietin (EPO). The results revealed that: (D In methylcellulose culture, the plating efficiencies of purified cord blood CD34+ cells were much different when stimulated by various cytokines. IL-6 alone had the lowest colo-ny yield, while the combination of SCF, IL-6 and EPO had the highest yield. ② In the suspension culture, IL-6 alone or IL-6 + EPO had little expanding effect on cord blood CD34+ celis, the other cytokine combinations could expand cord blood CD34+ celis at different Ievels. Among them, the combination of SCF, IL-6 and EPO had the maximal expanding effect on cord blood CD34+ celis, the number of progenitor celis peaked at day 21, about 29-fold increase and nucleated celis increased approximately 3676-fold at day 28. The expanding effect of 展开更多
关键词 cord blood cd34+ cell CYTOKINE ex vivo EXPANSION
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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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The effect of the different labeling and red cell removed methods on measuring the CD34^+cells of the cord blood
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《中国输血杂志》 CAS CSCD 2001年第S1期415-,共1页
关键词 cells of the cord blood cd The effect of the different labeling and red cell removed methods on measuring the cd34
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Experimental study on the expansion and hypothermic freezing of umbilical cord blood CD34^+ cells
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《中国输血杂志》 CAS CSCD 2001年第S1期417-,共1页
关键词 cd Experimental study on the expansion and hypothermic freezing of umbilical cord blood cd34
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The Cross-talk between ROS and p38MAPKα in the Ex Vivo Expanded Human Umbilical Cord Blood CD133^+ Cells 被引量:1
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作者 邹菁 邹萍 +3 位作者 罗毅 肖音 汪洁 刘凌波 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2011年第5期591-595,共5页
This study investigated the correlation between and compared the effects of reactive oxygen species(ROS) and p38 mitogen-activated protein kinase α(p38MAPKα) in the ex vivo expanded umbilical cord blood(hUCB) ... This study investigated the correlation between and compared the effects of reactive oxygen species(ROS) and p38 mitogen-activated protein kinase α(p38MAPKα) in the ex vivo expanded umbilical cord blood(hUCB) CD133+ cells.hUCB CD133+ cells were cultured in the hematopoietic stem cells(HSCs) culture medium with N-acetylcysteine(NAC,an anti-oxidant),p38MAPKα-specific inhibitor(SB203580) or their combination.The levels of ROS and expression of phosphorylated p38MAPKα(p-p38) in CD133+ cells were flow cytometrically detected.The efficacy of ex vivo expansion was evaluated by the density of CD133+ cell sub-group colony-forming cells(CFC) and cobblestone area-forming cells(CAFC) assay.Our results showed decreased ROS levels in NAC,SB203580,and their combination treatment groups were almost 37%,48%,and 85%,respectively.Furthermore,SB203580 abrogated the activation of p38MAPKα more obviously than NAC.Moreover,the CD133+ cells in SB203580 treatment group had a 21.93±1.36-fold increase,and 14.50±1.19-fold increase in NAC treatment group,but only 10.13±0.57-fold increase in control group.In addition,SB203580 treatment led a higher level increase in the number of CFU and CAFC than NAC did.These findings suggested that,in expanded CD133+ cells,ROS activates p38MAPKα,which,in turn,induces ROS production,and p38MAPKα might be the most suitable regulator in ROS-p38MAPKα pathway for the promotion of HSCs ex vivo expansion. 展开更多
关键词 p38 mitogen-activated protein kinase α reactive oxygen species human cord blood cd133+ cells hematopoiesis ex vivo expansion
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PERIPHERAL BLOOD CD34^+ CELL MOBILIZATION IN 42 PATIENTS WITH SEVERE AUTOIMMUNE DISEASE
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作者 Wei Zhang Dao-bin Zhou +8 位作者 Yan Zhao Jun-ling Zhuang Xiao-mei Leng Shu-jie Wang Li Jiao Fu-lin Tang Jie-ping Zhang Xuan Wang Ti Shen 《Chinese Medical Sciences Journal》 CAS CSCD 2007年第2期108-112,共5页
Objective To evaluate the feasibility and safety of peripheral CD34+ cell mobilization in patients with severe autoimmune disease. Methods Forty-two patients underwent a total of 46 mobilizations by the regimen of cyc... Objective To evaluate the feasibility and safety of peripheral CD34+ cell mobilization in patients with severe autoimmune disease. Methods Forty-two patients underwent a total of 46 mobilizations by the regimen of cyclophosphamide 2-3 g/m2 +recombinant human granulocyte colony stimulating factor (rhG-CSF) 5 μg·kg-1·d-1. The positive selection of CD34+ cell was performed through the CliniMACS. Results In 8.1±2.3 days after administration of cyclophosphamide, the peripheral white blood cell and mononuclear cell (MNC) decreased to the lowest level. In 3.7±1.6 days after injection of rhG-CSF, the peripheral absolute MNC and CD34+ cell counts were 0.95×109/L and 0.035×109/L, respectively. After 2.4±0.6 times of leukapheresis, there gained 4.46×108/kg of MNC and 5.26×106/kg of CD34+, respectively. After mobilization, the underlying diseases were ameliorated more or less. In systemic lupus erythematosus (SLE) patients, SLE Disease Activity Index (SLEDAI) decreased from a median of 17 to 3 (P<0.01). In rheumatic arthritis patients, an American College of Rheumatology criteria for 20%(ACR20) response was achieved in all five patients. Totally, 17.4% of patients whose absolute neutrophil count <0.5×109/L suffered infection, and 31.0% of patients had bone pain after the injection of rhG-CSF. Two patients suffered severe complications, one with acute renal failure and recovered by hemodialysis, the other died of thrombotic thrombocytopenic purpura. Failed mobilization occurred in three patients. Conclusions Sufficient CD34+ cells can be mobilized by low dose of cyclophosphamide and rhG-CSF. CD34+ cell mobilization for treatment of severe autoimmune disease not only is appropriate in both effectiveness and safety but ameliorates disease also. 展开更多
关键词 autoimmune disease ^cd34^+ cell mobilization
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Repetitive administration of cultured human CD34+cells improve adenine-induced kidney injury in mice
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作者 Takayasu Ohtake Shoichi Itaba +9 位作者 Amankeldi A Salybekov Yin Sheng Tsutomu Sato Mitsuru Yanai Makoto Imagawa Shigeo Fujii Hiroki Kumagai Masamitsu Harata Takayuki Asahara Shuzo Kobayashi 《World Journal of Stem Cells》 SCIE 2023年第4期268-280,共13页
BACKGROUND There is no established treatment to impede the progression or restore kidney function in human chronic kidney disease(CKD).AIM To examine the efficacy of cultured human CD34+cells with enhanced proliferati... BACKGROUND There is no established treatment to impede the progression or restore kidney function in human chronic kidney disease(CKD).AIM To examine the efficacy of cultured human CD34+cells with enhanced proliferating potential in kidney injury in mice.METHODS Human umbilical cord blood(UCB)-derived CD34+cells were incubated for one week in vasculogenic conditioning medium.Vasculogenic culture significantly increased the number of CD34+cells and their ability to form endothelial progenitor cell colony-forming units.Adenineinduced tubulointerstitial injury of the kidney was induced in immunodeficient non-obese diabetic/severe combined immunodeficiency mice,and cultured human UCB-CD34+cells were administered at a dose of 1×106/mouse on days 7,14,and 21 after the start of adenine diet.RESULTS Repetitive administration of cultured UCB-CD34+cells significantly improved the time-course of kidney dysfunction in the cell therapy group compared with that in the control group.Both interstitial fibrosis and tubular damage were significantly reduced in the cell therapy group compared with those in the control group(P<0.01).Microvasculature integrity was significantly preserved(P<0.01)and macrophage infiltration into kidney tissue was dramatically decreased in the cell therapy group compared with those in the control group(P<0.001).CONCLUSION Early intervention using human cultured CD34+cells significantly improved the progression of tubulointerstitial kidney injury.Repetitive administration of cultured human UCB-CD34+cells significantly improved tubulointerstitial damage in adenine-induced kidney injury in mice via vasculoprotective and anti-inflammatory effects. 展开更多
关键词 Chronic kidney disease cd34+cell ADENINE Tubulointerstitial injury Quality and quantity control culture Umbilical cord blood
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妊娠及围产因素对子代脐带血CD34^(+)造血干细胞的影响分析
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作者 仝晓鑫 李惠敏 +2 位作者 翟青梅 张璐涵 丁桂凤 《生物医学工程与临床》 CAS 2023年第6期759-763,共5页
目的分析妊娠及围产因素对子代CD34^(+)造血干细胞数量和脐带血有核细胞总数的影响。方法选择乌鲁木齐市妇幼保健院2021年5月至2022年5月收集的120例健康产妇的新生儿脐带血标本,其中男性60例,女性60例;新生儿体质量2500~4000 g。采用XE... 目的分析妊娠及围产因素对子代CD34^(+)造血干细胞数量和脐带血有核细胞总数的影响。方法选择乌鲁木齐市妇幼保健院2021年5月至2022年5月收集的120例健康产妇的新生儿脐带血标本,其中男性60例,女性60例;新生儿体质量2500~4000 g。采用XE-2100全自动血液分析仪对子代脐带血中的有核细胞进行计数。采用流式细胞分析仪分析CD34^(+)造血干细胞。收集新生儿及相应产妇的临床资料,主要包括孕母年龄、孕母血型、孕母身体质量指数(BMI)、孕母过敏史、妊娠期糖尿病、妊娠期高血压、分娩方式、产次、孕周、新生儿性别、新生儿体质量、胎盘质量、是否胎膜早破、是否羊水污染、是否脐带绕颈等信息,分析其对脐带血CD34^(+)造血干细胞影响。结果妊娠期因素主要有孕母年龄、孕母血型、孕母BMI、孕母过敏史、妊娠期糖尿病、妊娠期高血压、分娩方式、产次和孕周。围产期因素主要包括新生儿性别、新生儿体质量、胎盘质量、胎膜早破、羊水污染、脐带绕颈。根据临床信息分组进行组间比较发现,孕母血型、孕母BMI、孕母过敏史、妊娠期糖尿病、妊娠期高血压、分娩方式、产次、新生儿性别、新生儿体质量、胎盘质量、是否胎膜早破、是否羊水污染、是否脐带绕颈等因素与脐带血有核细胞总数和CD34^(+)造血干细胞数量无相关性(P>0.05),而孕母年龄和孕周均与CD34^(+)造血干细胞数量呈正相关(P<0.05)。结论子代脐带血中CD34^(+)造血干细胞数量与孕母年龄和孕周密切相关,孕母的年龄越大或孕周越大则CD34^(+)造血干细胞数量越多,这为脐血库筛选脐带血入库时提供了更多的理论依据。 展开更多
关键词 妊娠因素 围产因素 子代脐带血 ^cd34^(+)造血干细胞
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Analysis of CD4^+CD25^+ Regulatory T Cells and Foxp3 mRNA in the Peripheral Blood of Patients with Asthma 被引量:15
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作者 薛克营 周咏明 +2 位作者 熊盛道 熊维宁 唐滔 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2007年第1期31-33,共3页
The changes of CD4^+CD25^+ regulatory T cells (CD4^+CD25^+ Treg) and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma were investigated in order to elucidate the possible role... The changes of CD4^+CD25^+ regulatory T cells (CD4^+CD25^+ Treg) and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma were investigated in order to elucidate the possible roles of CD4^+CD25^+ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls (normal control group) and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^+CD25^+ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^+CD25^+ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups (P〈0.05). Although the CD4^+CD25^+ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them (P〉0.05). As compared with persistent group, exacerbation group had lower CD4^+CD25^+ Treg ratio and Foxp3 mRNA (P〈0.05). It was indicated that the decrease of CD4^+CD25^+ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma. 展开更多
关键词 ASTHMA peripheral blood mononuclear cells ^cd4^+cd25^+ regulatory T cells Foxp3 mRNA
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Functional expression of CD95/Fas Antigen and Bcl-2 on Cord blood Hematopoietic Progenitor Cells
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作者 MA Yanping(马艳萍) +2 位作者 ZOU Ping (邹萍) 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2002年第1期24-27,共4页
The cell surface expression and functional status of the CD95/Fas antigen on primitive hematopoietic progenitors isolated from human cord blood (CB) were studied. The CD34 + cells freshly isolated from CB displayed ... The cell surface expression and functional status of the CD95/Fas antigen on primitive hematopoietic progenitors isolated from human cord blood (CB) were studied. The CD34 + cells freshly isolated from CB displayed low CD95 expression. The combinations of cytokines such as SCF+FL could up regulate the expression of CD95 in vitro culture and tumor necrosis factor α (TNF α) and interon γ (IFN γ) further increased the CD95 expression induced by positive cytokines. The functional status of CD95 mediated apoptosis were analyzed by incubation of CD34 +CB cells in the presence of anti CD95 monoclonal antibodies (McAbs). The effects of anti CD95 McAbs were measured by viable cell counting, flow cytometry, LTIC and CFU C assays. A decrease of viable cells, CFU C and LTIC numbers were observed in the presence of anti CD95 McAbs and TNF α or IFN γ. However, growth factor deprivation or the early acting cytokine such as SCF and FL cross linking to CD95 caused low apoptosis of CD34 + cells. The correlation of increased intracytoplasmic levels of bcl 2 and the presence of CD95 on fresh CB CD34 + cells suggested that bcl 2 might be involved in protecting against CD95 mediated apoptosis of CB CD34 + cells. 展开更多
关键词 cd95/Fas Bcl 2 expression cord blood cd34 + cells
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Effect of Homoharringtonine on Bone Morrow CD34^+CD7^+ Cells in Chronic Granulocytic Leukemia
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作者 李玉峰 邓之奎 +1 位作者 宣衡报 陈宝安 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2007年第2期141-145,共5页
Objective: To explore the effect of homoharringtonine (HHT) on bone morrow CD34^+CD7^+ cells in chronic granulocytic leukemia (CGL). Methods: The changes of bone morrow CD34^+CD7^+ cells were observed after ... Objective: To explore the effect of homoharringtonine (HHT) on bone morrow CD34^+CD7^+ cells in chronic granulocytic leukemia (CGL). Methods: The changes of bone morrow CD34^+CD7^+ cells were observed after the treatment of HHT in 23 cases with CGL. The proliferation and apoptosis of CD34^+CD7^+ cells treated with HHT in vitro were studied. Results: The proportion of CD34^+CD7^+ cells in CGL (0.145±0.021) was higher than that of normal control (0.052±0.013). The proportion of CD34^+CD7^+ cells in patients who got cytogenetic responses to HHT (0.072±0.020) decreased remarkably, but not in those patients who did not got cytogenetic responses to HHT, (0.137±0.023). the proliferation of CD34^+ cells was inhibited and the proportion of CD34^+CD7^+ cells decreased after cultured with HHT (0.134 in 24 h, 0.126 in 48 h and 0.102 in 72). The apoptosis rate of CD34^+CD7^+ cells was higher than that in CD34^+CDT cells (35.39%±4.39% versus 24.57%±4.01%, P〈0.05) 72 h after culture with HHT. Conclusion: The proportion of CD34^+CD7^+ cells in CGL was higher than that of normal control and HHT may inhibit the proliferation and induce apoptosis of bone marrow CD34^+CD7^+ cells. 展开更多
关键词 HOMOHARRINGTONINE Chronic granulocytic leukemia ^cd34^+cd7^+ cells
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人胎盘源贴壁细胞支持脐血CD34^+细胞体外扩增 被引量:8
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作者 张毅 何津 +4 位作者 江小霞 刘刚 刘元林 唐佩弦 毛宁 《中国实验血液学杂志》 CAS CSCD 2003年第6期560-564,共5页
从胎盘中分离培养人胎盘源贴壁细胞 (humanplacentaderivedadherentcells,hPDAC) ,研究其对脐血CD34+细胞体外扩增作用。以酶消化法自人胎盘组织中分离培养hPDAC ,并以流式细胞术对其进行鉴定。进一步 ,采用免疫磁珠法分离人脐血CD34+细... 从胎盘中分离培养人胎盘源贴壁细胞 (humanplacentaderivedadherentcells,hPDAC) ,研究其对脐血CD34+细胞体外扩增作用。以酶消化法自人胎盘组织中分离培养hPDAC ,并以流式细胞术对其进行鉴定。进一步 ,采用免疫磁珠法分离人脐血CD34+细胞 ,建立以hPDAC为滋养层的CD34+细胞体外扩增培养体系 ,并与无滋养层的液体培养体系相比 ,观察不同培养体系对有核细胞总数、CFC及CD34+细胞百分率的影响。结果表明 ,人胎盘组织中可分离培养出hPDAC ,并证实其为混合性细胞群体 ,主要含有间充质细胞。以hPDAC为滋养层的共培养体系较无滋养层液体培养体系对有核细胞总数、CFC及CD34+细胞均具有明显的扩增效应 ,其中以SCF +IL 3+IL 6 +FL +hPDAC组扩增效果最强 ,分别扩增了 ( 12 6 .0± 6 .7)倍 ,( 4 9.8± 1.7)倍和 ( 8.3± 1.6 5 )倍。上述结果提示 ,hPDAC具有体外支持造血作用 ,并提供了一与脐血CD34+细胞体外扩增相适应的新滋养层。 展开更多
关键词 人胎盘源贴壁细胞 脐血 ^cd34^+细胞 hPDAC 生物学 细胞因子 抗体
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不同时相移植人骨髓间充质干细胞对人脐血CD34^+细胞植入NOD/SCID小鼠的影响 被引量:13
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作者 马俐君 胡晓霞 +3 位作者 周虹 高磊 邱慧颖 王健民 《中国实验血液学杂志》 CAS CSCD 2008年第2期355-359,共5页
为了观察不同时相移植人骨髓间充质干细胞(MSC)对脐血(UCB)CD34+细胞移植的NOD/SCID小鼠造血重建的影响,明确最佳的移植时机,将体外培养扩增的人骨髓MSC分别于c细胞移植同时、移植前48小时及移植后48小时输入经60Coγ射线照射的NOD/SCI... 为了观察不同时相移植人骨髓间充质干细胞(MSC)对脐血(UCB)CD34+细胞移植的NOD/SCID小鼠造血重建的影响,明确最佳的移植时机,将体外培养扩增的人骨髓MSC分别于c细胞移植同时、移植前48小时及移植后48小时输入经60Coγ射线照射的NOD/SCID小鼠,观察共移植后42天内小鼠外周血白细胞和血小板变化,并于移植后42天处死小鼠,用FACS检测外周血、骨髓和脾脏人源细胞含量。结果表明:(1)MSC和UCB CD34+细胞同时输注可明显降低外周血白细胞和血小板下降幅度,缩短白细胞和血小板恢复时间;二者不同时输注均不降低白细胞和血小板下降幅度,且输注UCB CD34+细胞后48小时输注MSC时外周血血小板恢复时间明显晚于同时输注者。(2)与单纯UCB CD34+细胞移植相比较,不同时相输注MSC均可促进UCB CD34+细胞的植入,三个共输注组间促进骨髓各系造血植入效应无明显差异。结论:人骨髓MSC与UCB CD34+细胞共移植时,以同时移植效果最佳,此结果为MSC的临床应用提供了实验依据。 展开更多
关键词 间充质干细胞 ^脐血cd34^+细胞 共移植 NOD/SCID小鼠
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GM-CSF对脐血CD34^+巨核祖细胞体外扩增及分化的影响 被引量:5
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作者 陈舒 朱发明 +3 位作者 何吉 刘晋辉 秦斐 严力行 《中国实验血液学杂志》 CAS CSCD 2005年第6期1041-1043,共3页
本实验旨在研究GMCSF对脐血CD34+细胞诱导分化为巨核细胞的影响。采用免疫磁珠法分选CD34+细胞,在含有TPO+IL3+SCF并添加了不同浓度(5、20、100ng/ml)的GMCSF的无血清培养基中进行培养。培养6、10、14天后计数单个核细胞(MNC),检测CD41... 本实验旨在研究GMCSF对脐血CD34+细胞诱导分化为巨核细胞的影响。采用免疫磁珠法分选CD34+细胞,在含有TPO+IL3+SCF并添加了不同浓度(5、20、100ng/ml)的GMCSF的无血清培养基中进行培养。培养6、10、14天后计数单个核细胞(MNC),检测CD41+细胞比例和CFUMK。结果表明,培养14天后3种不同浓度GMCSF对MNC均有明显的扩增作用,其中以20和100ng/mlGMCSF的扩增效果较好。3种不同浓度的GMCSF均使CD41+细胞比例增加,20和100ng/ml与5ng/mlGMCSF相比更能提高CD41+细胞的比例。5和20ng/ml的GMCSF能促进CFUMK的形成,但100ng/ml的GMCSF却抑制CFUMK的形成。结论:在TPO+IL3+SCF细胞因子组合中添加GMCSF有利于促进脐血CD34+细胞诱导分化为巨核细胞。 展开更多
关键词 GM-CSF 脐血 ^cd34^+细胞 巨核细胞 体外扩增
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人骨髓间充质干细胞体外支持脐血CD34^+细胞增殖的研究 被引量:7
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作者 马俐君 高磊 +4 位作者 周虹 邱慧颖 胡晓霞 解琳娜 王健民 《中国实验血液学杂志》 CAS CSCD 2006年第5期949-954,共6页
为了研究人骨髓间充质干细胞(MSC)作为滋养层细胞体外支持脐血CD34+细胞扩增的作用,将MSC和胎儿骨髓来源的成纤维细胞样骨髓基质细胞系(HFCL)经60Coγ线照射后分别制备细胞滋养层,比较不同滋养层细胞和添加或不加细胞因子对脐血CD34+细... 为了研究人骨髓间充质干细胞(MSC)作为滋养层细胞体外支持脐血CD34+细胞扩增的作用,将MSC和胎儿骨髓来源的成纤维细胞样骨髓基质细胞系(HFCL)经60Coγ线照射后分别制备细胞滋养层,比较不同滋养层细胞和添加或不加细胞因子对脐血CD34+细胞增殖数及LTC-IC数的影响。结果表明,无论有无添加细胞因子(rhFL、rhSCF、rhTPO),HFCL滋养层组培养12天扩增细胞数明显高于MSC滋养层组,以第0天细胞数为100%(下同),有细胞因子组为(9797±361)%vs(7061±418)%,无细胞因子组为(5305±354)%vs(1992±247)%,均P<0.01。MSC滋养层组CD34+细胞扩增数与HFCL滋养层组相比无显著差异[(825±305)%vs(820±191)%,P>0.05],但在细胞因子存在时低于HFCL滋养层组[(939±212)%vs(1617±222)%,P<0.01]。MSC滋养层组维持脐血中LTC-IC的能力明显优于HFCL滋养层组[第5周CFU-GM数(129.95±8.73)个/105接种细胞数vs(89.81±10.29)个/105接种细胞数,P<0.05];细胞因子存在时,其作用更为明显[第5周CFU-GM数(192.93±4.95)个/105接种细胞数vs(90.47±14.28)个/105接种细胞数,P<0.01]。MSC与HFCL按一定比例混合,可提高扩增效率。当MSC与HFCL之比为4∶1时,集落形成数量最高,达(186.89±11.11)个/105接种细胞数,明显高于比例为3∶2(131.45±13.02)个/105接种细胞数和二者单用组[前者(138.92±14.84)个/105接种细胞数,后者(64.63±6.11)个/105接种细胞数;均P<0.01]。结论:MSC维持脐血中LTC-IC集落形成的能力优于基质细胞系HFCL,HFCL支持脐血CD34+细胞的增殖能力优于MSC,MSC加上适量HFCL可显著提高CD34+细胞扩增效率。 展开更多
关键词 间充质干细胞 ^脐血cd34^+细胞 ^cd34^+细胞扩增
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脐血CD34^+细胞体外扩增时HOXB4基因表达的变化 被引量:6
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作者 唐宇宏 费小明 +3 位作者 沈文怡 缪扣荣 崔毓桂 汪承亚 《中国实验血液学杂志》 CAS CSCD 2006年第1期89-93,共5页
同源盒基因家族成员HOXB4反映原始造血干/祖细胞(PHSC/PHPC)的自我更新和增殖能力。本研究采用实时定量RT-PCR的方法在mRNA水平上检测HOXB4基因的表达,以观察体外扩增脐血CD34+细胞自我更新的水平。结果显示:随着体外培养时间的延长,虽... 同源盒基因家族成员HOXB4反映原始造血干/祖细胞(PHSC/PHPC)的自我更新和增殖能力。本研究采用实时定量RT-PCR的方法在mRNA水平上检测HOXB4基因的表达,以观察体外扩增脐血CD34+细胞自我更新的水平。结果显示:随着体外培养时间的延长,虽然CD34+细胞数增加,但HOXB4表达下降;周后,HOXB4几乎检测不到,与成熟外周血淋巴细胞表达HOXB4的水平相同;CD34+细胞与骨髓间充质干细胞(BM-MSC)共培养可以减缓CD34+细胞HOXB4表达的下降。结论:脐血CD34+细胞体外扩增过程中自我更新能力逐渐下降。CD34+与BM-MSC共培养有助于减缓体外扩增的CD34+细胞自我更新能力的丧失。 展开更多
关键词 HOXB4 ^脐血cd34^+细胞 体外扩增 骨髓间充质干细胞 实时定量RT—PCR
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人脐带血CD34^+细胞体外诱导树突状细胞及其激活T细胞抗肿瘤免疫反应的实验研究 被引量:2
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作者 王冠军 解丽华 +3 位作者 王晓 冯凯 李梁 裴雪涛 《中国实验血液学杂志》 CAS CSCD 2001年第2期188-190,共3页
To induce the growth and differentiation of dendritic cells (DCs) from human cord blood, CD34 + cells isolated from human cord blood by mini MACS were cultured in a liquid culture system with rhSCF, rhGM CSF, rhTNF α... To induce the growth and differentiation of dendritic cells (DCs) from human cord blood, CD34 + cells isolated from human cord blood by mini MACS were cultured in a liquid culture system with rhSCF, rhGM CSF, rhTNF α and rhFL for 10 days. Then the induced cells were characterized by DC′s morphological and phenotypic properties. In addition, they stimulated the proliferation of allogeneic T cells and possessed an efficient capacity for initiating T cell dependent antitumor immune responses in vitro. It is concluded that mature DCs could be obtained from human cord blood CD34 + cells. 展开更多
关键词 脐带血 ^cd34^+细胞 树突状细胞 T细胞 抗肿瘤免疫反应 实验研究
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细胞因子体外诱导脐血CD34^+细胞增殖并向巨核细胞/血小板分化的研究 被引量:2
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作者 张可莹 刘江 +7 位作者 贾延军 李伟 段澜 高颂明 崔爽 龚治尹 倪雷 张志欣 《中国实验血液学杂志》 CAS CSCD 2011年第4期1053-1057,共5页
本研究旨在观察几种不同细胞因子组合通过体外培养以诱导造血干/祖细胞增殖和向巨核细胞/血小板分化。应用无血清培养基(S tem Span(SFEM)体外扩增脐血CD34+细胞并向巨核细胞/血小板定向分化,将不同细胞因子组合分为3个阶段培养,并比较... 本研究旨在观察几种不同细胞因子组合通过体外培养以诱导造血干/祖细胞增殖和向巨核细胞/血小板分化。应用无血清培养基(S tem Span(SFEM)体外扩增脐血CD34+细胞并向巨核细胞/血小板定向分化,将不同细胞因子组合分为3个阶段培养,并比较其培养效果。结果表明,在第一阶段的第14天时,SCF+TPO+FL+IL-3组细胞扩增倍数最高为11 000±1 000,显著高于SCF+TPO+FL组,但与SCF+TPO+IL-3组及SCF+TPO+FL+IL-3+羟皮质激素组相比较无显著差异;在第二培养阶段的第7天时,SCF+TPO+FL+IL-11组巨核细胞系扩增倍数为204666.7±11718.9,显著高于SCF+TPO+FL+IL-3组,与SCF+TPO+FL+IL1-1+BM P4+VEGF组比较并无显著差异。在第三阶段中,SCF+TPO+FL+IL-11和SCF+TPO+FL+IL-11+BM P4+VEGF2组的CD41+血小板样细胞所占比例显著高于SCF+TPO+FL+IL-3组。结论:SCF+TPO+FL+IL-3因子组合可显著提高造血干/祖细胞的扩增倍数,SCF+TPO+FL+IL-11组合有利于巨核细胞的扩增成熟及分化,为下一步的体外培养巨核细胞/血小板体系的建立奠定了一定的基础。 展开更多
关键词 细胞因子 体外培养 血小板 脐血cd34’细胞 血小板增殖 血小板分化
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体外共培养双份脐血CD34^+细胞对各自归巢相关分子表达的影响 被引量:2
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作者 姚雯 汪健 +3 位作者 孙自敏 刘会兰 耿良权 王兴兵 《中国实验血液学杂志》 CAS CSCD 2008年第2期368-372,共5页
本研究探讨脐血CD34+细胞体外混合培养对彼此归巢相关分子表达的影响。从正常人骨髓中分离扩增间充质干细胞,富集传代,加速器照射(12Gy)后作为滋养层细胞,将免疫磁珠分选纯化的两份脐血CD34+细胞混合接种到其表面(其中1份CD34+细胞用CFS... 本研究探讨脐血CD34+细胞体外混合培养对彼此归巢相关分子表达的影响。从正常人骨髓中分离扩增间充质干细胞,富集传代,加速器照射(12Gy)后作为滋养层细胞,将免疫磁珠分选纯化的两份脐血CD34+细胞混合接种到其表面(其中1份CD34+细胞用CFSE标记),观察各自归巢相关分子(黏附分子)表达的变化。结果表明,脐血CD34+细胞分选富集的纯度为(98.25±0.93)%,实验组在共培养6天后,两份脐血CD34+细胞的比例分别降为(60.4±6.32)%和(60.2±5.12)%,但与对照组比较无统计学差异;实验组各份CD34+细胞的CD44,CD62L,CD184以及CD26的阳性率较培养前均无显著变化。而CD162表达显著下降。CD54在培养3天后有所上升,但培养6天后下降至培养前水平,与对照组比较无统计学差异。结论:将两份脐血的CD34+细胞体外混合培养对彼此归巢相关分子并无明显影响。 展开更多
关键词 脐血 ^cd34^+细胞 体外共培养 归巢
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