Browsing by Author "Chokdee Wongborisuth"

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    In vitro production of functional immune cells derived from human hematopoietic stem cells
    (Leibniz Research Centre for Working Environment and Human Factors, 2015) Witchuda Payuhakrit; Tasanee Panichakul; Natthawut Charoenphon; Panus Chalermsaenyakorn; Adithep Jaovisidha; Chokdee Wongborisuth; Rachanee Udomsangpetch
    Hematopoietic stem cells (HSC) from cord blood are potentially high sources for transplantation due to their low immunogenicity and the presence of the multipotent cells. These cells are capable of differentiating to produce various lineages of blood cells under specific conditions. We have enriched highly purified CD34+ cells from cord blood, determined in vitro growth of the cells in culture systems in the absence (condition A) or presence of GM-CSF and G-CSF (condition B), and determined the profile of immune cells during the period of cultivation by using flow cytometry. PhytohemagglutininA (PHA) was used as a mitogen to stimulate T lymphocytes derived from hematopoietic stem cells. GM-CSF and G-CSF prolonged the survival of the growing cells and also maintained expansion of cells in blastic stage. By day 12 of cultivation, when cell numbers peaked, various types of immune cells had appeared (CD14+ cells, CD40+HLA-DR+ cells, CD3+CD56+ cells, CD19+ cells, CD3+CD4+ cells, CD3+CD8+cells and CD3-CD56+). A significantly higher percentage of monocytes (p = 0.002) were observed under culture with GM-CSF, G-CSF when compared with culture without GM-CSF, G-CSF. In addition, T lymphocytes derived from HSC responded to 50 _g/ml of PHA. This is the first report showing the complete differentiation and proliferation of immune cells derived from CD34+ HSC under in vitro culture conditions. Lymphocytes, monocytes, dendritic cells and polymorph nuclear cells derived from HSC in vitro are unique, and thus may benefit various studies such as innate immunity and pathophysiology of immune disorders. � 2015, Leibniz Research Centre for Working Environment and Human Factors. All right reserved.
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    Phosphoproteomic analysis of apoptotic hematopoietic stem cells from hemoglobin E/_-thalassemia
    (2011) Saranyoo Ponnikorn; Tasanee Panichakul; Kitima Sresanga; Chokdee Wongborisuth; Sittiruk Roytrakul; Suradej Hongeng; Sumalee Tungpradabkul; S. Hongeng; Department of Pediatrics, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand; email: rashe@mahidol.ac.th
    Background: Hemoglobin E/_-thalassemia is particularly common in Southeast Asia and has variable symptoms ranging from mild to severe anemia. Previous investigations demonstrated the remarkable symptoms of _-thalassemia in terms of the acceleration of apoptotic cell death. Ineffective erythropoiesis has been studied in human hematopoietic stem cells, however the distinct apoptotic mechanism was unclear.Methods: The phosphoproteome of bone marrow HSCs/CD34+ cells from HbE/_-thalassemic patients was analyzed using IMAC phosphoprotein isolation followed by LC-MS/MS detection. Decyder MS software was used to quantitate differentially expressed proteins in 3 patients and 2 normal donors. The differentially expressed proteins from HSCs/CD34+ cells were compared with HbE/_-thalassemia and normal HSCs.Results: A significant change in abundance of 229 phosphoproteins was demonstrated. Importantly, the analysis of the candidate proteins revealed a high abundance of proteins that are commonly found in apoptotic cells including cytochrome C, caspase 6 and apoptosis inducing factors. Moreover, in the HSCs patients a significant increase was observed in a specific type of phosphoserine/threonine binding protein, which is known to act as an important signal mediator for the regulation of cell survival and apoptosis in HbE/_-thalassemia.Conclusions: Our study used a novel method to investigate proteins that influence a particular pathway in a given disease or physiological condition. Ultimately, phosphoproteome profiling in HbE/_-thalassemic stem cells is an effective method to further investigate the cell death mechanism of ineffective erythropoiesis in _-thalassemia. Our report provides a comprehensive phosphoproteome, an important resource for the study of ineffective erythropoiesis and developing therapies for HbE/_-thalassemia. © 2011 Ponnikorn et al; licensee BioMed Central Ltd.
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    Suppression of erythroid development in vitro by Plasmodium vivax
    (2012) Tasanee Panichakul; Witchuda Payuhakrit; Panyu Panburana; Chokdee Wongborisuth; Suradej Hongeng; Rachanee Udomsangpetch; T. Panichakul; Faculty of Science and Technology, Suan Dusit Rajabhat University, Bangplat, Bangkok, 10700, 204/3 Sirindhorn Rd., Thailand; email: tasanee_p@yahoo.com
    Background: Severe anaemia due to dyserythropoiesis has been documented in patients infected with Plasmodium vivax, however the mechanism responsible for anaemia in vivax malaria is poorly understood. In order to better understand the role of P. vivax infection in anaemia the inhibition of erythropoiesis using haematopoietic stem cells was investigated. Methods: Haematopoietic stem cells/CD34 + cells, isolated from normal human cord blood were used to generate growing erythroid cells. Exposure of CD34 + cells and growing erythroid cells to P. vivax parasites either from intact or lysed infected erythrocytes (IE) was examined for the effect on inhibition of cell development compared with untreated controls. Results: Both lysed and intact infected erythrocytes significantly inhibited erythroid growth. The reduction of erythroid growth did not differ significantly between exposure to intact and lysed IE and the mean growth relative to unexposed controls was 59.4 ± 5.2 for lysed IE and 57 ± 8.5% for intact IE. Interestingly, CD34 + cells/erythroid progenitor cells were susceptible to the inhibitory effect of P. vivax on cell expansion. Exposure to P. vivax also inhibited erythroid development, as determined by the reduced expression of glycophorin A (28.1%) and CD 71 (43.9%). Moreover, vivax parasites perturbed the division of erythroid cells, as measured by the Cytokinesis Block Proliferation Index, which was reduced to 1.35 ± 0.05 (P-value²0.01) from a value of 2.08 ± 0.07 in controls. Neither TNF-a nor IFN-g was detected in the culture medium of erythroid cells treated with P. vivax, indicating that impaired erythropoiesis was independent of these cytokines. Conclusions: This study shows for the first time that P. vivax parasites inhibit erythroid development leading to ineffective erythropoiesis and highlights the potential of P. vivax to cause severe anaemia. © 2012 Panichakul et al.; licensee BioMed Central Ltd.