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
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
为动态观察并探讨不同损伤程度的颅脑损伤病人外周血白细胞及CD_(34)^+细胞的变化及同预后的关系,以及二者升高的机制和临床意义,选取我院神经外科收治的伤后6h内入院脑外伤病人,按入院时格拉斯哥昏迷评分(Glasgow coma sale,GCS)分为...为动态观察并探讨不同损伤程度的颅脑损伤病人外周血白细胞及CD_(34)^+细胞的变化及同预后的关系,以及二者升高的机制和临床意义,选取我院神经外科收治的伤后6h内入院脑外伤病人,按入院时格拉斯哥昏迷评分(Glasgow coma sale,GCS)分为轻重两组,同时选择我院体检中心健康成年人15名,作为对照组。于入院第1天、第3天、第5天、第7天、第11天、第14天抽取静脉血,用流式细胞仪检测外周血CD_(34)^+细胞含量;用血细胞分析仪常规检测白细胞计数,同时观察病情变化,追踪预后,按照病人预后不同情况分成死亡组和生存组,比较分析相关数据。结果表明,30例颅脑损伤病人外周血白细胞及CD_(34)_^+细胞数量随病程的发展均有不同程度的升高,损伤越重、GCS评分越低,白细胞及CD_(34)^+细胞计数越高,预后越差,白细胞升高持续时间越长,预后愈差。外周血CD_(34)^+细胞在颅脑损伤后轻、重两组病人,各时间段逐步升高。重症病人自第3天与正常对照组出现统计学差异,明显高于轻症组病人,轻、重症两组间比较,在第7、11、14天有统计学差异。白细胞计数在伤后迅速升高并逐步下降,同CD_(34)^+细胞变化未见正相关。本研究认为,外周血白细胞及CD_(34)^+细胞,在颅脑损伤病人急性期均有不同程度的升高,计数增高越明显,持续时间越长,结果预后越差。外周血白细胞及CD_(34)^+细胞计数的变化可以成为判断其预后的重要指标。展开更多
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
Summary： The characteristics for the ex vivo expansion of the endothelial progenitor cells （EPCs） were explored, CD34^＋ cells were selected from umbilical cord blood mononuclear cells （MNC） by MiniMACS system, e...Summary： The characteristics for the ex vivo expansion of the endothelial progenitor cells （EPCs） were explored, CD34^＋ cells were selected from umbilical cord blood mononuclear cells （MNC） by MiniMACS system, expanded under the same conditions as those for total MNC, coincubation of CD34^＋ and CD34 from the same donor for EPCs. In addition, the effects of vessel endothelial growth factor （VEGF） and passage on cell differentiation, expansion kinetics and apoptosis were examined, EPCs were determined and quantified by immunocytochemistry and flow cytometry, The results showed that both coculture of CD346＋ and CD34^- and total MNC led to a significant increase in the expansion of CD34^＋ cells as compared with CD34 enrichment （P〈0.05）. There was a tendency toward decreased apoptosis in cultures when early passage was performed immediately after cord like structures appeared. VEGF had no significant effect on apoptosis （P〉0.05）, These differentiated EPCs were positive for CD34^＋, von Willebrand factor （vWF）, KDR, CD31 staining and phagocytized acetylated low-density lipoprotein （LDL）. CD34^＋ cells accounted for （68.2±6,3） % of attaching （AT） cells at day 7 of culture. It was suggested the most efficient method to ex vivo expansion of EPCs was coculture of CD34^＋ and CD34^- or total MNC. Early passage makes cell apoptosis rate decrease. VEGF had no significant effect on ex vivo expansion of EPCs.展开更多
Objective:To comprehensively explore hematopoietic stem cells(HSCs)in human milk,understanding their molecular markers,isolation methods,benefits for infants,and potential medical applications.Methods:We conducted a s...Objective:To comprehensively explore hematopoietic stem cells(HSCs)in human milk,understanding their molecular markers,isolation methods,benefits for infants,and potential medical applications.Methods:We conducted a scoping literature review following the PRISMA-ScR guidelines.This review included studies investigating HSCs in human milk,utilizing molecular markers such as CD34^(+),CD113^(+),and CD117^(+)for characterization.Both in vitro and in vivo studies exploring the morphology,function,and clinical implications of these cells were considered.The diverse range of papers reviewed were indexed in PubMed,Science Direct,Scopus,Sage Journals,and Google Scholar,published between 2010 and 2023.Results:This scoping review explored 577 articles and selected 13 studies based on our inclusion criteria,focusing on HSCs in human milk.Most studies dilute samples prior to HSC isolation,followed by detection using markers such as CD34^(+),CD113^(+),and CD117^(+),with flow cytometry serving as the primary analysis tool,focusing on their isolation and detection methods.While no definitive benefits have been conclusively established,there is a strong belief in the potential of HSCs to positively impact infant immunity,growth,and tissue repair.Conclusions:This review presents significant evidence supporting the presence of HSCs in human milk,identified by markers such as CD34^(+),CD113^(+),and CD117^(+).These cells show considerable potential in enhancing infant health,including immunity,tissue repair,cognitive development,and gastrointestinal health.Despite methodological variations in isolation and detection techniques,the collective findings underscore the potential clinical relevance of HSCs in human milk.Moreover,this review highlights the noninvasive accessibility of human milk as a source of HSCs and emphasizes the need for further research to unlock their therapeutic potential.展开更多
As a source of transplantable stem cells, the CD34^＋ subpopulation in human umbilical cord blood （HUCB） has been used extensively to treat some hematopoietic system diseases. However, whether CD34^＋ cells hold th...As a source of transplantable stem cells, the CD34^＋ subpopulation in human umbilical cord blood （HUCB） has been used extensively to treat some hematopoietic system diseases. However, whether CD34^＋ cells hold the therapeutic potential to cerebral ischemia is unknown. The purpose of this study was to observe the recovery of neural function after transplantation of CD34^＋ cells derived from HUCB into ischemic cerebral tissue in rats.展开更多
Objective To successfully transduce a mutant erbB 2 gene into normal human CD34 + derived dendritic cells (DCs) and verify gene expression in these transduced dendritic cells Methods The packaging cell line PA31...Objective To successfully transduce a mutant erbB 2 gene into normal human CD34 + derived dendritic cells (DCs) and verify gene expression in these transduced dendritic cells Methods The packaging cell line PA317 was transfected with mutant erbB 2 gene DNA and the virus produced was used to infect packaging cell line PG13 The virus produced by PG13 was used to infect CD34 + derived dendritic cells by the spinoculation method using flasks coated with fibronectin material to facilitate retrovirus gene transter efficiency and the mutant erbB 2 gene expression was assessed by ABC staining and FACScan methods Results A mutant erbB 2 gene packaging cell line was produced and this mutant gene was transduced into human CD34 + derived DCs It was verified that the relatively large numbers of the transduced DCs expressed the mutant erbB 2 protein which was eradicated of the ability to transform mouse NIH3T3 fibroblast cells Conclusions Human DCs can be gene modified and these gene modified DCs may be useful in stimulating T lymphocytes for immunotherapy展开更多
基金The study was supported by a grant from the National Natural Science Foundation of China(No.39928010)
文摘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.
文摘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.
文摘为动态观察并探讨不同损伤程度的颅脑损伤病人外周血白细胞及CD_(34)^+细胞的变化及同预后的关系,以及二者升高的机制和临床意义,选取我院神经外科收治的伤后6h内入院脑外伤病人,按入院时格拉斯哥昏迷评分(Glasgow coma sale,GCS)分为轻重两组,同时选择我院体检中心健康成年人15名,作为对照组。于入院第1天、第3天、第5天、第7天、第11天、第14天抽取静脉血,用流式细胞仪检测外周血CD_(34)^+细胞含量;用血细胞分析仪常规检测白细胞计数,同时观察病情变化,追踪预后,按照病人预后不同情况分成死亡组和生存组,比较分析相关数据。结果表明,30例颅脑损伤病人外周血白细胞及CD_(34)_^+细胞数量随病程的发展均有不同程度的升高,损伤越重、GCS评分越低,白细胞及CD_(34)^+细胞计数越高,预后越差,白细胞升高持续时间越长,预后愈差。外周血CD_(34)^+细胞在颅脑损伤后轻、重两组病人,各时间段逐步升高。重症病人自第3天与正常对照组出现统计学差异,明显高于轻症组病人,轻、重症两组间比较,在第7、11、14天有统计学差异。白细胞计数在伤后迅速升高并逐步下降,同CD_(34)^+细胞变化未见正相关。本研究认为,外周血白细胞及CD_(34)^+细胞,在颅脑损伤病人急性期均有不同程度的升高,计数增高越明显,持续时间越长,结果预后越差。外周血白细胞及CD_(34)^+细胞计数的变化可以成为判断其预后的重要指标。
文摘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.
基金This project was supported by a grant from Hubei Provin-cial Bureau of Health (No. WI01532).
文摘Summary： The characteristics for the ex vivo expansion of the endothelial progenitor cells （EPCs） were explored, CD34^＋ cells were selected from umbilical cord blood mononuclear cells （MNC） by MiniMACS system, expanded under the same conditions as those for total MNC, coincubation of CD34^＋ and CD34 from the same donor for EPCs. In addition, the effects of vessel endothelial growth factor （VEGF） and passage on cell differentiation, expansion kinetics and apoptosis were examined, EPCs were determined and quantified by immunocytochemistry and flow cytometry, The results showed that both coculture of CD346＋ and CD34^- and total MNC led to a significant increase in the expansion of CD34^＋ cells as compared with CD34 enrichment （P〈0.05）. There was a tendency toward decreased apoptosis in cultures when early passage was performed immediately after cord like structures appeared. VEGF had no significant effect on apoptosis （P〉0.05）, These differentiated EPCs were positive for CD34^＋, von Willebrand factor （vWF）, KDR, CD31 staining and phagocytized acetylated low-density lipoprotein （LDL）. CD34^＋ cells accounted for （68.2±6,3） % of attaching （AT） cells at day 7 of culture. It was suggested the most efficient method to ex vivo expansion of EPCs was coculture of CD34^＋ and CD34^- or total MNC. Early passage makes cell apoptosis rate decrease. VEGF had no significant effect on ex vivo expansion of EPCs.
基金supported by the National Research and Innovation Agency of Republic of Indonesia(BRIN)-RIIM Batch-22022 research grants and the Institute of Education Fund Management(Lembaga Pengelola Dana Pendidikan-LPDP).
文摘Objective:To comprehensively explore hematopoietic stem cells(HSCs)in human milk,understanding their molecular markers,isolation methods,benefits for infants,and potential medical applications.Methods:We conducted a scoping literature review following the PRISMA-ScR guidelines.This review included studies investigating HSCs in human milk,utilizing molecular markers such as CD34^(+),CD113^(+),and CD117^(+)for characterization.Both in vitro and in vivo studies exploring the morphology,function,and clinical implications of these cells were considered.The diverse range of papers reviewed were indexed in PubMed,Science Direct,Scopus,Sage Journals,and Google Scholar,published between 2010 and 2023.Results:This scoping review explored 577 articles and selected 13 studies based on our inclusion criteria,focusing on HSCs in human milk.Most studies dilute samples prior to HSC isolation,followed by detection using markers such as CD34^(+),CD113^(+),and CD117^(+),with flow cytometry serving as the primary analysis tool,focusing on their isolation and detection methods.While no definitive benefits have been conclusively established,there is a strong belief in the potential of HSCs to positively impact infant immunity,growth,and tissue repair.Conclusions:This review presents significant evidence supporting the presence of HSCs in human milk,identified by markers such as CD34^(+),CD113^(+),and CD117^(+).These cells show considerable potential in enhancing infant health,including immunity,tissue repair,cognitive development,and gastrointestinal health.Despite methodological variations in isolation and detection techniques,the collective findings underscore the potential clinical relevance of HSCs in human milk.Moreover,this review highlights the noninvasive accessibility of human milk as a source of HSCs and emphasizes the need for further research to unlock their therapeutic potential.
基金This work was supported by a grant from the Natural Science Foundation of Hebei Province (No. 302508).
文摘As a source of transplantable stem cells, the CD34^＋ subpopulation in human umbilical cord blood （HUCB） has been used extensively to treat some hematopoietic system diseases. However, whether CD34^＋ cells hold the therapeutic potential to cerebral ischemia is unknown. The purpose of this study was to observe the recovery of neural function after transplantation of CD34^＋ cells derived from HUCB into ischemic cerebral tissue in rats.
文摘Objective To successfully transduce a mutant erbB 2 gene into normal human CD34 + derived dendritic cells (DCs) and verify gene expression in these transduced dendritic cells Methods The packaging cell line PA317 was transfected with mutant erbB 2 gene DNA and the virus produced was used to infect packaging cell line PG13 The virus produced by PG13 was used to infect CD34 + derived dendritic cells by the spinoculation method using flasks coated with fibronectin material to facilitate retrovirus gene transter efficiency and the mutant erbB 2 gene expression was assessed by ABC staining and FACScan methods Results A mutant erbB 2 gene packaging cell line was produced and this mutant gene was transduced into human CD34 + derived DCs It was verified that the relatively large numbers of the transduced DCs expressed the mutant erbB 2 protein which was eradicated of the ability to transform mouse NIH3T3 fibroblast cells Conclusions Human DCs can be gene modified and these gene modified DCs may be useful in stimulating T lymphocytes for immunotherapy
