The Experts below are selected from a list of 1824174 Experts worldwide ranked by ideXlab platform
Hiroyuki Suzuki - One of the best experts on this subject based on the ideXlab platform.
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evaluation of the skin sensitization potential of chemicals using expression of co stimulatory molecules cd54 and cd86 on the naive thp 1 cell line
Toxicology in Vitro, 2003Co-Authors: Yukiko Yoshida, Hitoshi Sakaguchi, Yuichi Ito, M Okuda, Hiroyuki SuzukiAbstract:It has been known that dendritic cells (DCs) including Langerhans cells (LCs) play a critical role in the skin sensitization process. Many attempts have been made to develop in vitro sensitization tests that employ DCs derived from peripheral blood mononuclear cells (PBMC-DC) or CD34+ hematopoietic progenitor cells (CD34+ HPC) purified from cord blood or bone marrow. However, the use of the DCs in in vitro methods has been difficult due to the nature of these cells such as low levels in the source and/or donor-to-donor variability. In our studies, we employed the human monocytic leukemia cell line, THP-1, in order to avoid some of these difficulties. At the start, we examined whether treatment of the cells with various cytokines could produce DCs from THP-1. Treatment of THP-1 cells with cytokines such as GM-CSF, IL-4, TNF-alpha, and/or PMA did induce some phenotypic changes in THP-1 cells that were characteristic of DCs. Subsequently, responses to a known sensitizer, dinitrochlorobenzene (DNCB), and a non-sensitizer, dimethyl sulfoxide (DMSO) or sodium lauryl sulfate (SLS), on the expression of co-stimulatory molecules, CD54 and CD86, were examined between the naive cells and the cytokine-treated cells. Interestingly, the naive THP-1 cells responded only to DNCB and the response to the sensitizer was more distinct than cytokine-treated THP-1 cells. Similar phenomena were also observed in the human myeloid leukemia cell line, KG-1. Furthermore, with treatment of DNCB, naive THP-1 cells showed augmented expression of HLA, CD80 and secretion of IL-1 beta. The response of THP-1 cells to a sensitizer was similar to that of LCs/DCs. Upon demonstrating the differentiation of monocyte cells in our system, we then evaluated a series of chemicals, including known sensitizers and non-sensitizers, for their potential to augment CD54 and CD86 expression on naive THP-1 cells. Indeed, known sensitizers such as PPD and 2-MBT significantly augmented CD54 and CD86 expression in a dose-dependent manner while non-sensitizers, such as SLS and methyl salicylate (MS), did not. To note, the metal allergens such as (NH(4))(2)[PtCl(4)], NiSO(4) and CoSO(4) augmented significantly only CD54 expression. Taking advantage of a cultured cell line, measurement of the co-stimulatory molecules, CD54 and CD86, on naive THP-1 cells following chemical exposure shows promise for the development of a simple, short-term in vitro sensitization test.
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evaluation of the skin sensitization potential of chemicals using expression of co stimulatory molecules cd54 and cd86 on the naive thp 1 cell line
Toxicology in Vitro, 2003Co-Authors: Yukiko Yoshida, Hitoshi Sakaguchi, M Okuda, Hiroyuki SuzukiAbstract:Abstract It has been known that dendritic cells (DCs) including Langerhans cells (LCs) play a critical role in the skin sensitization process. Many attempts have been made to develop in vitro sensitization tests that employ DCs derived from peripheral blood mononuclear cells (PBMC-DC) or CD34+ hematopoietic progenitor cells (CD34+ HPC) purified from cord blood or bone marrow. However, the use of the DCs in in vitro methods has been difficult due to the nature of these cells such as low levels in the source and/or donor-to-donor variability. In our studies, we employed the human monocytic leukemia cell line, THP-1, in order to avoid some of these difficulties. At the start, we examined whether treatment of the cells with various cytokines could produce DCs from THP-1. Treatment of THP-1 cells with cytokines such as GM-CSF, IL-4, TNF-α, and/or PMA did induce some phenotypic changes in THP-1 cells that were characteristic of DCs. Subsequently, responses to a known sensitizer, dinitrochlorobenzene (DNCB), and a non-sensitizer, dimethyl sulfoxide (DMSO) or sodium lauryl sulfate (SLS), on the expression of co-stimulatory molecules, CD54 and CD86, were examined between the naive cells and the cytokine-treated cells. Interestingly, the naive THP-1 cells responded only to DNCB and the response to the sensitizer was more distinct than cytokine-treated THP-1 cells. Similar phenomena were also observed in the human myeloid leukemia cell line, KG-1. Furthermore, with treatment of DNCB, naive THP-1 cells showed augmented expression of HLA, CD80 and secretion of IL-1β. The response of THP-1 cells to a sensitizer was similar to that of LCs/DCs. Upon demonstrating the differentiation of monocyte cells in our system, we then evaluated a series of chemicals, including known sensitizers and non-sensitizers, for their potential to augment CD54 and CD86 expression on naive THP-1 cells. Indeed, known sensitizers such as PPD and 2-MBT significantly augmented CD54 and CD86 expression in a dose-dependent manner while non-sensitizers, such as SLS and methyl salicylate (MS), did not. To note, the metal allergens such as (NH4)2[PtCl4], NiSO4 and CoSO4 augmented significantly only CD54 expression. Taking advantage of a cultured cell line, measurement of the co-stimulatory molecules, CD54 and CD86, on naive THP-1 cells following chemical exposure shows promise for the development of a simple, short-term in vitro sensitization test.
Yukiko Yoshida - One of the best experts on this subject based on the ideXlab platform.
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evaluation of the skin sensitization potential of chemicals using expression of co stimulatory molecules cd54 and cd86 on the naive thp 1 cell line
Toxicology in Vitro, 2003Co-Authors: Yukiko Yoshida, Hitoshi Sakaguchi, Yuichi Ito, M Okuda, Hiroyuki SuzukiAbstract:It has been known that dendritic cells (DCs) including Langerhans cells (LCs) play a critical role in the skin sensitization process. Many attempts have been made to develop in vitro sensitization tests that employ DCs derived from peripheral blood mononuclear cells (PBMC-DC) or CD34+ hematopoietic progenitor cells (CD34+ HPC) purified from cord blood or bone marrow. However, the use of the DCs in in vitro methods has been difficult due to the nature of these cells such as low levels in the source and/or donor-to-donor variability. In our studies, we employed the human monocytic leukemia cell line, THP-1, in order to avoid some of these difficulties. At the start, we examined whether treatment of the cells with various cytokines could produce DCs from THP-1. Treatment of THP-1 cells with cytokines such as GM-CSF, IL-4, TNF-alpha, and/or PMA did induce some phenotypic changes in THP-1 cells that were characteristic of DCs. Subsequently, responses to a known sensitizer, dinitrochlorobenzene (DNCB), and a non-sensitizer, dimethyl sulfoxide (DMSO) or sodium lauryl sulfate (SLS), on the expression of co-stimulatory molecules, CD54 and CD86, were examined between the naive cells and the cytokine-treated cells. Interestingly, the naive THP-1 cells responded only to DNCB and the response to the sensitizer was more distinct than cytokine-treated THP-1 cells. Similar phenomena were also observed in the human myeloid leukemia cell line, KG-1. Furthermore, with treatment of DNCB, naive THP-1 cells showed augmented expression of HLA, CD80 and secretion of IL-1 beta. The response of THP-1 cells to a sensitizer was similar to that of LCs/DCs. Upon demonstrating the differentiation of monocyte cells in our system, we then evaluated a series of chemicals, including known sensitizers and non-sensitizers, for their potential to augment CD54 and CD86 expression on naive THP-1 cells. Indeed, known sensitizers such as PPD and 2-MBT significantly augmented CD54 and CD86 expression in a dose-dependent manner while non-sensitizers, such as SLS and methyl salicylate (MS), did not. To note, the metal allergens such as (NH(4))(2)[PtCl(4)], NiSO(4) and CoSO(4) augmented significantly only CD54 expression. Taking advantage of a cultured cell line, measurement of the co-stimulatory molecules, CD54 and CD86, on naive THP-1 cells following chemical exposure shows promise for the development of a simple, short-term in vitro sensitization test.
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evaluation of the skin sensitization potential of chemicals using expression of co stimulatory molecules cd54 and cd86 on the naive thp 1 cell line
Toxicology in Vitro, 2003Co-Authors: Yukiko Yoshida, Hitoshi Sakaguchi, M Okuda, Hiroyuki SuzukiAbstract:Abstract It has been known that dendritic cells (DCs) including Langerhans cells (LCs) play a critical role in the skin sensitization process. Many attempts have been made to develop in vitro sensitization tests that employ DCs derived from peripheral blood mononuclear cells (PBMC-DC) or CD34+ hematopoietic progenitor cells (CD34+ HPC) purified from cord blood or bone marrow. However, the use of the DCs in in vitro methods has been difficult due to the nature of these cells such as low levels in the source and/or donor-to-donor variability. In our studies, we employed the human monocytic leukemia cell line, THP-1, in order to avoid some of these difficulties. At the start, we examined whether treatment of the cells with various cytokines could produce DCs from THP-1. Treatment of THP-1 cells with cytokines such as GM-CSF, IL-4, TNF-α, and/or PMA did induce some phenotypic changes in THP-1 cells that were characteristic of DCs. Subsequently, responses to a known sensitizer, dinitrochlorobenzene (DNCB), and a non-sensitizer, dimethyl sulfoxide (DMSO) or sodium lauryl sulfate (SLS), on the expression of co-stimulatory molecules, CD54 and CD86, were examined between the naive cells and the cytokine-treated cells. Interestingly, the naive THP-1 cells responded only to DNCB and the response to the sensitizer was more distinct than cytokine-treated THP-1 cells. Similar phenomena were also observed in the human myeloid leukemia cell line, KG-1. Furthermore, with treatment of DNCB, naive THP-1 cells showed augmented expression of HLA, CD80 and secretion of IL-1β. The response of THP-1 cells to a sensitizer was similar to that of LCs/DCs. Upon demonstrating the differentiation of monocyte cells in our system, we then evaluated a series of chemicals, including known sensitizers and non-sensitizers, for their potential to augment CD54 and CD86 expression on naive THP-1 cells. Indeed, known sensitizers such as PPD and 2-MBT significantly augmented CD54 and CD86 expression in a dose-dependent manner while non-sensitizers, such as SLS and methyl salicylate (MS), did not. To note, the metal allergens such as (NH4)2[PtCl4], NiSO4 and CoSO4 augmented significantly only CD54 expression. Taking advantage of a cultured cell line, measurement of the co-stimulatory molecules, CD54 and CD86, on naive THP-1 cells following chemical exposure shows promise for the development of a simple, short-term in vitro sensitization test.
Mason A Israel - One of the best experts on this subject based on the ideXlab platform.
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cell surface marker signatures for the isolation of neural stem cells glia and neurons derived from human pluripotent stem cells
PLOS ONE, 2011Co-Authors: Shauna H Yuan, Jody L Martin, Jeanne Elia, Jessica Flippin, Rosanto I Paramban, Michael P Hefferan, Jason G Vidal, Rhiannon L Killian, Mason A Israel, Nil EmreAbstract:Background Neural induction of human pluripotent stem cells often yields heterogeneous cell populations that can hamper quantitative and comparative analyses. There is a need for improved differentiation and enrichment procedures that generate highly pure populations of neural stem cells (NSC), glia and neurons. One way to address this problem is to identify cell-surface signatures that enable the isolation of these cell types from heterogeneous cell populations by fluorescence activated cell sorting (FACS). Methodology/Principal Findings We performed an unbiased FACS- and image-based immunophenotyping analysis using 190 antibodies to cell surface markers on naive human embryonic stem cells (hESC) and cell derivatives from neural differentiation cultures. From this analysis we identified prospective cell surface signatures for the isolation of NSC, glia and neurons. We isolated a population of NSC that was CD184+/CD271−/CD44−/CD24+ from neural induction cultures of hESC and human induced pluripotent stem cells (hiPSC). Sorted NSC could be propagated for many passages and could differentiate to mixed cultures of neurons and glia in vitro and in vivo. A population of neurons that was CD184−/CD44−/CD15LOW/CD24+ and a population of glia that was CD184+/CD44+ were subsequently purified from cultures of differentiating NSC. Purified neurons were viable, expressed mature and subtype-specific neuronal markers, and could fire action potentials. Purified glia were mitotic and could mature to GFAP-expressing astrocytes in vitro and in vivo. Conclusions/Significance These findings illustrate the utility of immunophenotyping screens for the identification of cell surface signatures of neural cells derived from human pluripotent stem cells. These signatures can be used for isolating highly pure populations of viable NSC, glia and neurons by FACS. The methods described here will enable downstream studies that require consistent and defined neural cell populations.
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cell surface marker signatures for the isolation of neural stem cells glia and neurons derived from human pluripotent stem cells
PLOS ONE, 2011Co-Authors: Shauna H Yuan, Jody L Martin, Jeanne Elia, Jessica Flippin, Rosanto I Paramban, Michael P Hefferan, Jason G Vidal, Rhiannon L Killian, Yangling Mu, Mason A IsraelAbstract:Background Neural induction of human pluripotent stem cells often yields heterogeneous cell populations that can hamper quantitative and comparative analyses. There is a need for improved differentiation and enrichment procedures that generate highly pure populations of neural stem cells (NSC), glia and neurons. One way to address this problem is to identify cell-surface signatures that enable the isolation of these cell types from heterogeneous cell populations by fluorescence activated cell sorting (FACS). Methodology/Principal Findings We performed an unbiased FACS- and image-based immunophenotyping analysis using 190 antibodies to cell surface markers on naive human embryonic stem cells (hESC) and cell derivatives from neural differentiation cultures. From this analysis we identified prospective cell surface signatures for the isolation of NSC, glia and neurons. We isolated a population of NSC that was CD184+/CD271−/CD44−/CD24+ from neural induction cultures of hESC and human induced pluripotent stem cells (hiPSC). Sorted NSC could be propagated for many passages and could differentiate to mixed cultures of neurons and glia in vitro and in vivo. A population of neurons that was CD184−/CD44−/CD15LOW/CD24+ and a population of glia that was CD184+/CD44+ were subsequently purified from cultures of differentiating NSC. Purified neurons were viable, expressed mature and subtype-specific neuronal markers, and could fire action potentials. Purified glia were mitotic and could mature to GFAP-expressing astrocytes in vitro and in vivo. Conclusions/Significance These findings illustrate the utility of immunophenotyping screens for the identification of cell surface signatures of neural cells derived from human pluripotent stem cells. These signatures can be used for isolating highly pure populations of viable NSC, glia and neurons by FACS. The methods described here will enable downstream studies that require consistent and defined neural cell populations.
Feng Wang - One of the best experts on this subject based on the ideXlab platform.
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a novel mouse model of human breast cancer stem like cells with high cd44 cd24 lower phenotype metastasis to human bone
Cancer Research, 2009Co-Authors: Lijun Ling, Shui Wang, Xiaoan Liu, Enchao Shen, Qiang Ding, Qinhong Cao, Hong Zhu, Feng WangAbstract:CTRC-AACR San Antonio Breast Cancer Symposium: 2008 Abstracts Abstract #4155 Background: At present, our understanding of the development of bone metastasis is limited. Therefore, little progress has been made in preventing skeletal metastasis in the breast cancer patient. A satisfactory animal model that avoids the species-specific factor and simultaneously shares similarities to the clinical pathophysiological progression of breast cancer metastasizing to bone is unavailable. A subpopulation (CD44+/CD24-/lower) of breast cancer cells possesses stem/progenitor cell properties (cancer stem-like cell). CD44 potentiates the adherence of metastatic breast cancer cells to bone marrow endothelial cells. In the present study, we used hepatocyte growth factor to enhance the proportion of CD44+/CD24-/lower subpopulation in the human breast cancer cell line MDA-MB-231. We used these human breast cancer stem-like cells and implantation of human bone to build a novel human-source model of human breast cancer skeletal metastasis. Methods: The human breast cancer cell line MDA-MB-231 was cultured in serum-free DMEM-F12 supplemented with growth factors. Cells in different subpopulations were separated by Percoll gradient centrifugation and incubated in the presence of 50 ng/mL hepatocyte growth factor. The proportion of CD44+/CD24-/lower subpopulation in the human breast cancer cell line MDA-MB-231 was detected with flow cytometry. Before injection with human breast cancer stem-like cells, the experimental animals were implanted with human bone in the right or left dorsal flanks. Animals in Groups A, B, and C were injected with 1 X 105, 1 X 106 human breast cancer stem-like cells, and 1 X 106 parental MDA-MB-231 cells, respectively. A positive control group (D) without implantation of human bone (non-human source model) was also injected with 1 X 106 MDA-MB-231 cells. A group of negative controls (E) with human bone implantation was injected with isotonic sodium chloride. For each group, Micro-SPECT was performed at weeks 4 and 7, and all animals were sacrificed at week 8. Immunohistochemistry was performed for CD34, CD105, SMA, CD44, CD24, CK, CXCR4, and OPN. mRNA levels of CD44, CD24, CXCR4, and OPN in bone metastasis tissues were analyzed by real-time quantitative polymerase chain reaction. Results: New vessels and connective tissue were found on the surface of implanted human bones and cells stained positive for antibodies against human CD105, SMA, and CD34, indicating that implanted human bones were viable and functional. Histologic and immunohistochemical analysis confirmed the metastases as cancer cells. Importantly, the results demonstrated that cells in implanted human bones of group B, which received 1 X 106 cancer stem-like cells, stained strongly positive for CD44, CXCR4, and OPN, whereas that of other groups showed no or minimum staining. Moreover, group B had the highest incidence of human bone metastasis (77.8%, P = 0.0230) and no accompaniment of other tissue metastasis. The real-time polymerase chain reaction (PCR) showed an increase of CD44 mRNA in metastatic bone tissues in group B compared with that of groups C and D (15.2- and 21.1-fold, respectively). The mRNA levels of CXCR4 and OPN (8.4- and 28.4-fold, respectively) in bone metastasis tissues of group B were all higher than that of groups C and D (4.8- and 11.6-fold; respectively). The levels of CD24 mRNA in group B were lowest, measuring only 30 percent of that in groups C and D. Conclusion: This study indicates that in the novel human source model of breast cancer, breast cancer stem-like cells demonstrate a higher human bone-seeking ability, which may contribute to increase metastasis incidence and attenuate species-specific influences. Its mechanism might be related to the higher expressions of CD44, CXCR4 and OPN, and the lower expression of CD24 in breast cancer stem-like cells. The model shares more similarities with clinical pathological features of bone metastatic patients. It will be helpful for further study of the mechanisms and subclinical diagnosis of bone metastasis. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 4155.
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a novel mouse model of human breast cancer stem like cells with high cd44 cd24 lower phenotype metastasis to human bone
Chinese Medical Journal, 2008Co-Authors: Lijun Ling, Shui Wang, Xiaoan Liu, Enchao Shen, Qiang Ding, Qinhong Cao, Hong Zhu, Feng WangAbstract:BACKGROUND A satisfactory animal model of breast cancer metastasizing to bone is unavailable. In this study, we used human breast cancer stem-like cells and human bone to build a novel "human-source" model of human breast cancer skeletal metastasis. METHODS Human breast cancer stem-like cells, the CD44+/CD24-/lower subpopulation, was separated and cultured. Before injection with the stem-like cells, mice were implanted with human bone in the right or left dorsal flanks. Animals in Groups A, B, and C were injected with 1 x 10(5), 1 x 10(6) human breast cancer stem-like cells, and 1 x 10(6) parental MDA-MB-231 cells, respectively. A positive control group (D) without implantation of human bone was also injected with 1 x 10(6) MDA-MB-231 cells. Immunohistochemistry was performed for determination of CD34, CD105, smooth muscle antibody, CD44, CD24, cytokine, CXC chemokine receptor-4 (CXCR4), and osteopontin (OPN). mRNA levels of CD44, CD24, CXCR4, and OPN in bone metastasis tissues were analyzed by real-time quantitative polymerase chain reaction (PCR). RESULTS Our results demonstrated that cells in implanted human bones of group B, which received 1 x 10(6) cancer stem-like cells, stained strongly positive for CD44, CXCR4, and OPN, whereas those of other groups showed no or minimum staining. Moreover, group B had the highest incidence of human bone metastasis (77.8%, P = 0.0230) and no accompaniment of other tissue metastasis. The real-time PCR showed an increase of CD44, CXCR4, and OPN mRNA in metastatic bone tissues in group B compared with those of groups C and D, however the expression of CD24 mRNA in group B were the lowest. CONCLUSIONS In the novel "human source" model of breast cancer, breast cancer stem-like cells demonstrated a higher human bone-seeking ability. Its mechanism might be related to the higher expressions of CD44, CXCR4, and OPN, and the lower expression of CD24 in breast cancer stem-like cells.
Shauna H Yuan - One of the best experts on this subject based on the ideXlab platform.
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cell surface marker signatures for the isolation of neural stem cells glia and neurons derived from human pluripotent stem cells
PLOS ONE, 2011Co-Authors: Shauna H Yuan, Jody L Martin, Jeanne Elia, Jessica Flippin, Rosanto I Paramban, Michael P Hefferan, Jason G Vidal, Rhiannon L Killian, Mason A Israel, Nil EmreAbstract:Background Neural induction of human pluripotent stem cells often yields heterogeneous cell populations that can hamper quantitative and comparative analyses. There is a need for improved differentiation and enrichment procedures that generate highly pure populations of neural stem cells (NSC), glia and neurons. One way to address this problem is to identify cell-surface signatures that enable the isolation of these cell types from heterogeneous cell populations by fluorescence activated cell sorting (FACS). Methodology/Principal Findings We performed an unbiased FACS- and image-based immunophenotyping analysis using 190 antibodies to cell surface markers on naive human embryonic stem cells (hESC) and cell derivatives from neural differentiation cultures. From this analysis we identified prospective cell surface signatures for the isolation of NSC, glia and neurons. We isolated a population of NSC that was CD184+/CD271−/CD44−/CD24+ from neural induction cultures of hESC and human induced pluripotent stem cells (hiPSC). Sorted NSC could be propagated for many passages and could differentiate to mixed cultures of neurons and glia in vitro and in vivo. A population of neurons that was CD184−/CD44−/CD15LOW/CD24+ and a population of glia that was CD184+/CD44+ were subsequently purified from cultures of differentiating NSC. Purified neurons were viable, expressed mature and subtype-specific neuronal markers, and could fire action potentials. Purified glia were mitotic and could mature to GFAP-expressing astrocytes in vitro and in vivo. Conclusions/Significance These findings illustrate the utility of immunophenotyping screens for the identification of cell surface signatures of neural cells derived from human pluripotent stem cells. These signatures can be used for isolating highly pure populations of viable NSC, glia and neurons by FACS. The methods described here will enable downstream studies that require consistent and defined neural cell populations.
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cell surface marker signatures for the isolation of neural stem cells glia and neurons derived from human pluripotent stem cells
PLOS ONE, 2011Co-Authors: Shauna H Yuan, Jody L Martin, Jeanne Elia, Jessica Flippin, Rosanto I Paramban, Michael P Hefferan, Jason G Vidal, Rhiannon L Killian, Yangling Mu, Mason A IsraelAbstract:Background Neural induction of human pluripotent stem cells often yields heterogeneous cell populations that can hamper quantitative and comparative analyses. There is a need for improved differentiation and enrichment procedures that generate highly pure populations of neural stem cells (NSC), glia and neurons. One way to address this problem is to identify cell-surface signatures that enable the isolation of these cell types from heterogeneous cell populations by fluorescence activated cell sorting (FACS). Methodology/Principal Findings We performed an unbiased FACS- and image-based immunophenotyping analysis using 190 antibodies to cell surface markers on naive human embryonic stem cells (hESC) and cell derivatives from neural differentiation cultures. From this analysis we identified prospective cell surface signatures for the isolation of NSC, glia and neurons. We isolated a population of NSC that was CD184+/CD271−/CD44−/CD24+ from neural induction cultures of hESC and human induced pluripotent stem cells (hiPSC). Sorted NSC could be propagated for many passages and could differentiate to mixed cultures of neurons and glia in vitro and in vivo. A population of neurons that was CD184−/CD44−/CD15LOW/CD24+ and a population of glia that was CD184+/CD44+ were subsequently purified from cultures of differentiating NSC. Purified neurons were viable, expressed mature and subtype-specific neuronal markers, and could fire action potentials. Purified glia were mitotic and could mature to GFAP-expressing astrocytes in vitro and in vivo. Conclusions/Significance These findings illustrate the utility of immunophenotyping screens for the identification of cell surface signatures of neural cells derived from human pluripotent stem cells. These signatures can be used for isolating highly pure populations of viable NSC, glia and neurons by FACS. The methods described here will enable downstream studies that require consistent and defined neural cell populations.