The Experts below are selected from a list of 56877 Experts worldwide ranked by ideXlab platform
Davide Sciuscio - One of the best experts on this subject based on the ideXlab platform.
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toward refinement of the colony forming Unit granulocyte macrophage clonogenic assay inclusion of a metabolic system
Toxicology in Vitro, 2006Co-Authors: Gianni Dal Negro, Luca Vandin, Monica Bonato, Davide SciuscioAbstract:This work represents a first attempt to refine the Colony-Forming Unit-granulocyte/macrophage (CFU-GM) clonogenic assay by incorporating liver microsomes and co-factors as a metabolic system into the in vitro test system in response to an ECVAM recommendation. From the comparison of results obtained with the CFU-GM clonogenic assay currently used and with the new experimental protocol, different toxicity on granulocyte/macrophage precursors was demonstrated, when drugs with a known metabolism in vivo were tested.
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a new experimental protocol as an alternative to the colony forming Unit granulocyte macrophage cfu gm clonogenic assay to assess the haematotoxic potential of new drugs
Toxicology in Vitro, 2006Co-Authors: Gianni Dal Negro, Luca Vandin, Monica Bonato, Paolo Repeto, Davide SciuscioAbstract:Abstract In this work, a first attempt to set-up a new in vitro experimental protocol with culture in liquid medium and flow cytometry analysis of bone marrow progenitors is described. This protocol is proposed as an alternative to the Colony-Forming Unit-granulocyte/macrophage (CFU-GM) clonogenic in vitro assay currently used to assess the toxic potential of new drugs in the bone marrow. This new experimental approach should enable to speed up the procedure of the in vitro haematotoxic potential assessment, to reduce inter-experimental variability and to enhance result accuracy. Preliminary results obtained demonstrated that the progenitor cell count by flow cytometry replacing the light microscopy granulocyte/macrophage colony count represents a tremendous improvement in terms of accuracy and standardisation. Moreover, differential counts of cell sub-populations can be performed by using specific monoclonal antibodies. Furthermore, this method demonstrated to be time-saving, since 4 day cell incubation period is required instead of 7–14 day incubation in the CFU-GM clonogenic assay. On the basis of results obtained so far, the new experimental protocol proposed looks a promising alternative to the CFU-GM clonogenic assay currently used.
Gianni Dal Negro - One of the best experts on this subject based on the ideXlab platform.
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toward refinement of the colony forming Unit granulocyte macrophage clonogenic assay inclusion of a metabolic system
Toxicology in Vitro, 2006Co-Authors: Gianni Dal Negro, Luca Vandin, Monica Bonato, Davide SciuscioAbstract:This work represents a first attempt to refine the Colony-Forming Unit-granulocyte/macrophage (CFU-GM) clonogenic assay by incorporating liver microsomes and co-factors as a metabolic system into the in vitro test system in response to an ECVAM recommendation. From the comparison of results obtained with the CFU-GM clonogenic assay currently used and with the new experimental protocol, different toxicity on granulocyte/macrophage precursors was demonstrated, when drugs with a known metabolism in vivo were tested.
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a new experimental protocol as an alternative to the colony forming Unit granulocyte macrophage cfu gm clonogenic assay to assess the haematotoxic potential of new drugs
Toxicology in Vitro, 2006Co-Authors: Gianni Dal Negro, Luca Vandin, Monica Bonato, Paolo Repeto, Davide SciuscioAbstract:Abstract In this work, a first attempt to set-up a new in vitro experimental protocol with culture in liquid medium and flow cytometry analysis of bone marrow progenitors is described. This protocol is proposed as an alternative to the Colony-Forming Unit-granulocyte/macrophage (CFU-GM) clonogenic in vitro assay currently used to assess the toxic potential of new drugs in the bone marrow. This new experimental approach should enable to speed up the procedure of the in vitro haematotoxic potential assessment, to reduce inter-experimental variability and to enhance result accuracy. Preliminary results obtained demonstrated that the progenitor cell count by flow cytometry replacing the light microscopy granulocyte/macrophage colony count represents a tremendous improvement in terms of accuracy and standardisation. Moreover, differential counts of cell sub-populations can be performed by using specific monoclonal antibodies. Furthermore, this method demonstrated to be time-saving, since 4 day cell incubation period is required instead of 7–14 day incubation in the CFU-GM clonogenic assay. On the basis of results obtained so far, the new experimental protocol proposed looks a promising alternative to the CFU-GM clonogenic assay currently used.
Monica Bonato - One of the best experts on this subject based on the ideXlab platform.
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toward refinement of the colony forming Unit granulocyte macrophage clonogenic assay inclusion of a metabolic system
Toxicology in Vitro, 2006Co-Authors: Gianni Dal Negro, Luca Vandin, Monica Bonato, Davide SciuscioAbstract:This work represents a first attempt to refine the Colony-Forming Unit-granulocyte/macrophage (CFU-GM) clonogenic assay by incorporating liver microsomes and co-factors as a metabolic system into the in vitro test system in response to an ECVAM recommendation. From the comparison of results obtained with the CFU-GM clonogenic assay currently used and with the new experimental protocol, different toxicity on granulocyte/macrophage precursors was demonstrated, when drugs with a known metabolism in vivo were tested.
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a new experimental protocol as an alternative to the colony forming Unit granulocyte macrophage cfu gm clonogenic assay to assess the haematotoxic potential of new drugs
Toxicology in Vitro, 2006Co-Authors: Gianni Dal Negro, Luca Vandin, Monica Bonato, Paolo Repeto, Davide SciuscioAbstract:Abstract In this work, a first attempt to set-up a new in vitro experimental protocol with culture in liquid medium and flow cytometry analysis of bone marrow progenitors is described. This protocol is proposed as an alternative to the Colony-Forming Unit-granulocyte/macrophage (CFU-GM) clonogenic in vitro assay currently used to assess the toxic potential of new drugs in the bone marrow. This new experimental approach should enable to speed up the procedure of the in vitro haematotoxic potential assessment, to reduce inter-experimental variability and to enhance result accuracy. Preliminary results obtained demonstrated that the progenitor cell count by flow cytometry replacing the light microscopy granulocyte/macrophage colony count represents a tremendous improvement in terms of accuracy and standardisation. Moreover, differential counts of cell sub-populations can be performed by using specific monoclonal antibodies. Furthermore, this method demonstrated to be time-saving, since 4 day cell incubation period is required instead of 7–14 day incubation in the CFU-GM clonogenic assay. On the basis of results obtained so far, the new experimental protocol proposed looks a promising alternative to the CFU-GM clonogenic assay currently used.
Luca Vandin - One of the best experts on this subject based on the ideXlab platform.
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toward refinement of the colony forming Unit granulocyte macrophage clonogenic assay inclusion of a metabolic system
Toxicology in Vitro, 2006Co-Authors: Gianni Dal Negro, Luca Vandin, Monica Bonato, Davide SciuscioAbstract:This work represents a first attempt to refine the Colony-Forming Unit-granulocyte/macrophage (CFU-GM) clonogenic assay by incorporating liver microsomes and co-factors as a metabolic system into the in vitro test system in response to an ECVAM recommendation. From the comparison of results obtained with the CFU-GM clonogenic assay currently used and with the new experimental protocol, different toxicity on granulocyte/macrophage precursors was demonstrated, when drugs with a known metabolism in vivo were tested.
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a new experimental protocol as an alternative to the colony forming Unit granulocyte macrophage cfu gm clonogenic assay to assess the haematotoxic potential of new drugs
Toxicology in Vitro, 2006Co-Authors: Gianni Dal Negro, Luca Vandin, Monica Bonato, Paolo Repeto, Davide SciuscioAbstract:Abstract In this work, a first attempt to set-up a new in vitro experimental protocol with culture in liquid medium and flow cytometry analysis of bone marrow progenitors is described. This protocol is proposed as an alternative to the Colony-Forming Unit-granulocyte/macrophage (CFU-GM) clonogenic in vitro assay currently used to assess the toxic potential of new drugs in the bone marrow. This new experimental approach should enable to speed up the procedure of the in vitro haematotoxic potential assessment, to reduce inter-experimental variability and to enhance result accuracy. Preliminary results obtained demonstrated that the progenitor cell count by flow cytometry replacing the light microscopy granulocyte/macrophage colony count represents a tremendous improvement in terms of accuracy and standardisation. Moreover, differential counts of cell sub-populations can be performed by using specific monoclonal antibodies. Furthermore, this method demonstrated to be time-saving, since 4 day cell incubation period is required instead of 7–14 day incubation in the CFU-GM clonogenic assay. On the basis of results obtained so far, the new experimental protocol proposed looks a promising alternative to the CFU-GM clonogenic assay currently used.
Hongli Zhang - One of the best experts on this subject based on the ideXlab platform.
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colony forming Unit cell cfu c assays at diagnosis cfu g m cluster predicts overall survival in myelodysplastic syndrome patients independently of ipss r
Oncotarget, 2016Co-Authors: Jinqin Liu, Robert Peter Gale, Ruixian Xing, Tiejun Qin, Yue Zhang, Liwei Fang, Zhen Song, J Q Liu, Yansong Ren, Hongli ZhangAbstract:// Bing Li 1,2,* , Jinqin Liu 2,* , Shiqiang Qu 1,* , Robert Peter Gale 3 , Zhen Song 4 , Ruixian Xing 1 , Junxia Liu 5 , Yansong Ren 5 , Zefeng Xu 1,2 , Tiejun Qin 1 , Yue Zhang 1,2 , Liwei Fang 1 , Hongli Zhang 1 , Lijuan Pan 1 , Naibo Hu 1 , Wenyu Cai 6 , Peihong Zhang 6 , Gang Huang 7 and Zhijian Xiao 1,2 1 MDS and MPN Centre, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China 2 State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China 3 Department of Medicine, Haematology Section, Division of Experimental Medicine, Imperial College, London, United Kingdom 4 Medical Service Division, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China 5 Cell Culture Laboratory, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China 6 Department of Pathology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China 7 Divisions of Experimental Hematology and Cancer Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, Ohio, USA * Bing Li, Jinqin Liu and Shiqiang Qu have contributed equally to this study Correspondence to: Zhijian Xiao, email: // Keywords : myelodysplastic syndromes; Colony-Forming Unit cell; prognosis Received : April 29, 2016 Accepted : August 25, 2016 Published : September 18, 2016 Abstract Background: In vitro Colony-Forming Unit cell (CFU-C) assays are usually-used to detect the quantitative and qualitative features of haematopoietic stem cells (HSCs). We studies CFU-C assays in bone marrow samples from 365 consecutive subjects with newly-diagnosed myelodysplastic syndrome (MDS). Data were interrogated for associations with prognosis. Methods: CFU-C assays were performed according to the protocol of MethoCultTM H4435 Enriched. 365 consecutive newly-diagnosed, untreated subjects with MDS diagnosed from July, 2007 to April, 2014 were studied. All subjects were reclassified according to the 2008 WHO criteria. Subjects were observed for survival until July 31, 2015. Follow-up data were available for 289 (80%) subjects. Median follow-up of survivors was 22 months (range, 1-85) months. Erythroid and myeloid colonies were isolated from each subject with one cytogenetic abnormality such as del(5/5q), +8, del(7/7q) or del(20q). Cytogenetic abnormalities of each colony were analyzed by fluorescence in situ hybridization (FISH). SPSS 17.0 software was used to make statistical analysis. Results: The numbers of burst-forming Units-erythroid (BFU-E), colony forming Unit-erythroid (CFU-E) and colony forming Unit-granulocytes/macrophages (CFU-G/M) were significantly lower than normals. A high ratio of cluster- to CFU-G/M was associated with poor-risk cytogenetics. In multivariable analyses a cluster- to CFU-G/M ratio >0.6 was an independent risk-factor for OS after adjusting for IPSS-R (HR 3.339, [95%CI 1.434-7.778]; P = 0.005) in very high-risk cohort. Conclusion: These data suggest abnormalities of proliferation and differentiation of erythroid and myeloid precursor cells in vitro parallel the ineffective hematopoiesis typical of MDS and may be useful in predicting outcomes of persons with higher-risk MDS.
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Colony-Forming Unit Cell (CFU-C) Assays in Myelodysplastic Syndrome
Blood, 2015Co-Authors: Jinqin Liu, Robert Peter Gale, Ruixian Xing, Tiejun Qin, Yue Zhang, Liwei Fang, Hongli ZhangAbstract:Introduction: The myelodysplastic syndromes (MDS) are a group of clonal diseases derived from hematopoietic stem cells (HSC). Colony-Forming Unit cell (CFU-C) assay is an effective method to study the number and the function of HSC in vitro. In this study, we focus on the characteristics and the prognostic value of CFU-C in patients with MDS. Patients and Method: CFU-C assays were performed according to the protocol of MethoCultTM H4435 Enriched (STEMCELL Technologies). A colony was defined as an aggregate of >40 cells. Clusters consisted of 4 to 40 cells. 560 consecutive newly-diagnosed, untreated subjects with MDS diagnosed from March, 2001 to April, 2013 were studied. All subjects were reclassified according to the 2008 WHO criteria. 535 subjects with evaluable cytogenetics were classified using the International Prognostic Scoring System (IPSS) and the revised International Prognostic Scoring System (IPSS-R) criteria. Follow-up data were available for 470£¨84%£©subjects. Median follow-up of survivors was 26 months (range, 1-170) months. Subjects receiving an allotransplants were censored in survival analyses. Erythroid and myeloid colonies were isolated from each subject with one cytogenetic abnormality such as del(5/5q-) or +8. Cytogenetic abnormalities of each colony were analyzed by fluorescence in situ hybridization (FISH). SPSS 17.0 software was used to make statistical analysis. Results: Frequencies of burst-forming Units-erythroid (BFU-E), colony forming Unit-erythroid (CFU-E) and colony forming Unit-granulocytes/macrophages (CFU-G/M) were significantly lower than normals (P 0.6) was associated with poor-risk cytogenetics (Table 2) and with worse overall survival in univariable (Figure 1, P=0.001) and multivariable analyses (HR 1.748, [1.01-3.0]; P=0.046) after adjusting for IPSS. Conclusions: These data suggest abnormalities of proliferation and differentiation of erythroid and myeloid precursor cells in vitro parallel the ineffective hematopoiesis typical of MDS and may be useful in predicting outcomes of patients with MDS. Disclosures No relevant conflicts of interest to declare.