Sasitorn Rungarunlert - Academia.edu (original) (raw)
Papers by Sasitorn Rungarunlert
World Journal of Stem Cells, 2009
Embryonic stem (ES) cells have the ability to differ entiate into all germ layers, holding great ... more Embryonic stem (ES) cells have the ability to differ entiate into all germ layers, holding great promise not only for a model of early embryonic development but also for a robust cell source for cellreplacement therapies and for drug screening. Embryoid body (EB) formation from ES cells is a common method for producing different cell lineages for further applications. However, conventional techniques such as hanging drop or static suspension culture are either inherently incapable of large scale production or exhibit limited control over cell aggregation during EB formation and subsequent EB aggregation. For standardized mass EB production, a well defined scale-up platform is necessary. Recently, novel scenario methods of EB formation in hydrodynamic conditions created by bioreactor culture systems using stirred suspension systems (spinner flasks), rotating cell culture system and rotary orbital culture have allowed largescale EB formation. Their use allows for continuous monitoring and control of the physical and chemical environment which is difficult to achieve by traditional methods. This review summarizes the current state of production of EBs derived from pluripotent cells in various culture systems. Furthermore, an overview of high quality EB formation strategies coupled with systems for in vitro differentiation into various cell types to be applied in cell replacement therapy is provided in this review. Recently, new insights in induced pluripotent stem (iPS) cell technology showed that differentiation and lineage commitment are not irreversible processes and this has opened new avenues in stem cell research. These cells are equivalent to ES cells in terms of both selfrenewal and differentiation capacity. Hence, culture systems for expansion and differentiation of iPS cells can also apply methodologies developed with ES cells, although direct evidence of their use for iPS cells is still limited.
Cellular reprogramming, 2012
Mouse embryonic stem cells (ESCs) and induced pluripotent stem (iPS) cells can be used as models ... more Mouse embryonic stem cells (ESCs) and induced pluripotent stem (iPS) cells can be used as models of neuronal differentiation for the investigation of mammalian neurogenesis, pharmacological testing, and development of cell-based therapies. Recently, mouse iPS cell lines have been generated by Sleeping Beauty (SB) transposon-mediated transgenesis (SB-iPS). In this study, we determined for the first time the differentiation potential of mouse SB-iPS cells to form neuronal progenitor cells (NPCs) and neurons. Undifferentiated SB-iPS and ES cells were aggregated into embryoid bodies (EBs) and cultured in neuronal differentiation medium supplemented with 5 μM all-trans retinoic acid. Thereafter, EBs were dissociated and plated to observe further neuronal differentiation. Samples were fixed on days 10 and 14 for immunocytochemistry staining using the NPC markers Pax6 and Nestin and the neuron marker βIII-tubulin/Tuj1. Nestin-labeled cells were analyzed further by flow cytometry. Our resul...
Journal of the Medical Association of Thailand = Chotmaihet thangphaet, 2008
To determine the effect of leukemia inhibitory factor (LIF) on the quality of in vitro produced m... more To determine the effect of leukemia inhibitory factor (LIF) on the quality of in vitro produced mouse blastocyst and the efficiency of embryonic stem (ES) cell derivation. Experimental study Reproductive Medicine Unit, Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University In vivo fertilized zygotes were collected and subjected to in vitro culture in potassium simplex optimized medium (KSOM) containing 1,000 unit/ml LIF. The developmental ability of the zygote to blastocyst-stage and the cell numbers in blastocysts were evaluated Expanded blastocysts developed in different culture media were subsequently subjected to ES cell derivation. MAIN OUTCOME MEASURE (s): The influence of LIF on the quality of and the total cell numbers of blastocyst developed in vitro. Supplementation of LIF in KSOM increased the rate of hatching blastocysts (63.8% vs. 53.7%; p < 0.05) and total cell numbers (91.4 +/- 15.0 vs. 85.1 +/- 7.7; p < 0.05) compared to KSOM alo...
ABSTRACT Regenerative cell therapy against cardiovascular disease would require mass production a... more ABSTRACT Regenerative cell therapy against cardiovascular disease would require mass production and purification of specific cell types before transplantation. To enable large-scale production of embryonic stem (ES)-derived pure cardiomyocytes, we developed an animal model for a single-step scalable bioprocess that allows direct embryoid body (EB) formation in a fully controlled slow-turning lateral vessel (STLV, Synthecon, Inc., Houston, TX, USA) bioreactor following inoculation with a single cell suspension of mouse ES cells. To enhance the yield of cardiac progenitor cells, mouse ES cells (HM1; 129Sv/Ola, Magin et al. 1992 Nucl. Acids Res. 20, 3795-3796) were targeted with the cardiac-specific mouse Nkx2.5 promoter driven enhanced fluorescent green protein (EGFP). Among 15 targeted colonies, which were characterised based on morphology, the ability to form EB, EGFP expression, and in vitro differentiation ability toward cardiomyocytes, 3 lines were further evaluated for the efficiency of cardiomyocyte production. The 3 lines were cultured in STLV bioreactor and compared with classical hanging drop (HD) and static suspension culture methods. Embryonic bodies at day 3 to 8 were collected and analysed by using fluorescence-activated cell sorting for markers of pluripotency (e.g. Oct-4, SSEA1, Nanog) and cardiac (e.g. Nkx2.5, Troponin T) lineage commitments. Data was analysed by one-way ANOVA and t-tests. The results showed that both level and kinetics of Nkx2.5 expression was dependent on culture conditions. The STLV and static suspension culture methods produced higher rates of Nkx2.5-positive cells on day 5 than that of HD (70 and 54 v. 30%, respectively). The STLV method produced a highly uniform population of efficiently differentiating EB in large quantities and resulted in the highest, 10(8) yield of cardiomyocytes in a single 110-mL STLV on day 4. In conclusion, the STLV method provides a technological platform for controlled large-scale generation of ES-cell-derived cardiomyocytes for clinical and industrial applications. In vivo transplantation tests of cardiomyocytes produced via STLV are currently underway.
The aims of this study were to investigate the meiotic competence of immature porcine oocytes fol... more The aims of this study were to investigate the meiotic competence of immature porcine oocytes following open pulled straw (OPS) vitrification and warming and also to test the efficiency of the pretreatment of immature oocytes with CB prior to OPS vitrification. Experiment 1, cumulus oocyte complexes (COCs) were randomly assigned to OPS vitrification, cryoprotectant (CPA) toxicity test and non-CPA/non-vitrified controls. Experiment 2, the COCs were treated with 7.5 μg/ml CB for 30 min. The oocytes were either immediately fixed or further incubated in a CB-free medium for 5 h, non-CB treated COCs served as controls. Experiment 3, CB-pretreated COCs were vitrified. The COCs exposed to CB and vitrification solutions and non-CPA, non-CB treated oocytes served as controls. In all cases, all the COCs were matured in vitro for 40-44 h. After IVM, the oocytes were examined for the stages of nuclear maturation. Vitrifying and warming significantly reduced the capability of the immature porcine oocytes to resume and reach metaphase (M II) (21.7%), compared unfavorably with the control . This, however, was not the result from the cryoprotactants per se, since the number of CPA-treated oocytes reaching MII stage did not significantly differ from the control (76% vs. 81.8%, p>0.05). While cytochalasin B depolymerized actin cytoskeleton, this effect was completely reversible during 5 h of culture. Pretreatment with CB before the vitrification of immature porcine oocytes significantly yielded greater MII rates compared with non-CB oocytes (34.9% vs. 21.7%). In conclusion, porcine immature oocytes can be cryopreserved by OPS vitrification but it adversely affects the meiotic competence. Cytochalasin B reversibly depolymerizes actin microfilaments and improves the cryopreservability of porcine immature oocytes.
Cellular Reprogramming, 2013
Embryonic stem cells (ESCs) have the ability to form aggregates, which are called embryoid bodies... more Embryonic stem cells (ESCs) have the ability to form aggregates, which are called embryoid bodies (EBs). EBs mimic early embryonic development and are commonly produced for cardiomyogenesis. Here, we describe a method of EB formation in hydrodynamic conditions using a slow-turning lateral vessel (STLV) bioreactor and the subsequent differentiation of EBs into cardiomyocytes. EBs formed in the STLV were compared with conventional techniques, such as hanging drop (HD) or static suspension cell culture (SSC), for homogeneity of EB size, shape, proliferation, apoptosis, and in vitro cardiac differentiation. After 3 days of culture, a four-fold improvement in the yield of EB formation/mL, a six-fold enhancement in total yield of EB/mL, and a nearly 10fold reduction of cells that failed to incorporate into EBs were achieved in STLV versus SSC. During cardiac differentiation, a 1.5-to 4.2-fold increase in the area of cardiac troponin T (cTnT) per single EB in STLV versus SSC and HD was achieved. These results demonstrate that the STLV method improves the quality and quantity of ES cells to form EBs and enhances the efficiency of cardiac differentiation. We have demonstrated that the mechanical method of cell differentiation creates different microenvironments for the cells and thus influences their lineage commitments, even when genetic origin and the culture medium are the same. Ascorbic acid (ASC) improved further cardiac commitment in differentiation assays. Hence, this culture system is suitable for the production of large numbers of cells for clinical cell replacement therapies and industrial drug testing applications.
Neuroscience Letters, 2014
• Inhibition of TGF-1/ALK pathway efficiently decrease contamination of pluripotent cells in ESC... more • Inhibition of TGF-1/ALK pathway efficiently decrease contamination of pluripotent cells in ESC-derived neuronal population.
World Journal of Stem Cells, 2009
Embryonic stem (ES) cells have the ability to differ entiate into all germ layers, holding great ... more Embryonic stem (ES) cells have the ability to differ entiate into all germ layers, holding great promise not only for a model of early embryonic development but also for a robust cell source for cellreplacement therapies and for drug screening. Embryoid body (EB) formation from ES cells is a common method for producing different cell lineages for further applications. However, conventional techniques such as hanging drop or static suspension culture are either inherently incapable of large scale production or exhibit limited control over cell aggregation during EB formation and subsequent EB aggregation. For standardized mass EB production, a well defined scale-up platform is necessary. Recently, novel scenario methods of EB formation in hydrodynamic conditions created by bioreactor culture systems using stirred suspension systems (spinner flasks), rotating cell culture system and rotary orbital culture have allowed largescale EB formation. Their use allows for continuous monitoring and control of the physical and chemical environment which is difficult to achieve by traditional methods. This review summarizes the current state of production of EBs derived from pluripotent cells in various culture systems. Furthermore, an overview of high quality EB formation strategies coupled with systems for in vitro differentiation into various cell types to be applied in cell replacement therapy is provided in this review. Recently, new insights in induced pluripotent stem (iPS) cell technology showed that differentiation and lineage commitment are not irreversible processes and this has opened new avenues in stem cell research. These cells are equivalent to ES cells in terms of both selfrenewal and differentiation capacity. Hence, culture systems for expansion and differentiation of iPS cells can also apply methodologies developed with ES cells, although direct evidence of their use for iPS cells is still limited.
Molecular Biotechnology, 2014
Embryonic stem cell (ESC)-derived cardiomyocytes are a promising cell source for the screening fo... more Embryonic stem cell (ESC)-derived cardiomyocytes are a promising cell source for the screening for potential cytoprotective molecules against ischemia/reperfusion injury, however, little is known on their behavior in hypoxia/reoxygenation conditions. Here we tested the cytoprotective effect of the NO-donor SNAP and its downstream cellular pathway. Mouse ESC-derived cardiomyocytes were subjected to 150-min simulated ischemia (SI) followed by 120-min reoxygenation or corresponding non-ischemic conditions. The following treatments were applied during SI or normoxia: the NO-donor S-Nitroso-N-acetyl-D,L-penicillamine (SNAP), the protein kinase G (PKG) inhibitor, the K ATP channel blocker glibenclamide, the particulate guanylate cyclase activator brain type natriuretic peptide (BNP), and a non-specific NO synthase inhibitor (N-Nitro-L-arginine, L-NNA) alone or in different combinations. Viability of cells was assayed by propidium iodide staining. SNAP attenuated SI-induced cell death in a concentration-dependent manner, and this protection was attenuated by inhibition of either PKG or K ATP channels. However, SI-induced cell death was not A. Görbe and Z. V. Varga have contributed equally to this study.
The Thai veterinary medicine
Derivation of embryonic stem (ES) cells from parthenogenetic embryos represents a possible altern... more Derivation of embryonic stem (ES) cells from parthenogenetic embryos represents a possible alternative approach to create histocompatible cells for regenerative medicine. The objectives of this study were to establish mouse parthenogenetic ES (pES) cell lines from parthenogenetically-derived blastocysts as a model system for human and animal research and to examine pluripotency differences among the pES cell lines. We are able to report the successful establishment of four pluripotent pES cell lines from blastocysts of parthenogenetic origin (22% efficiency of pES cell line establishment). Four pES cell lines (pES#1-4) exhibited a typical ES cell morphology and expression of key pluripotency markers (ALP, Oct4, Nanog and SSEA-1). Three of the four pES cell lines have shown a high percentage of normal karyotype during long-term culture. Variability in the in vitro differentiation potential into cell types of the 3 germ layers was observed among the different pES cell lines. Three of ...
World Journal of Stem Cells, 2009
Embryonic stem (ES) cells have the ability to differ entiate into all germ layers, holding great ... more Embryonic stem (ES) cells have the ability to differ entiate into all germ layers, holding great promise not only for a model of early embryonic development but also for a robust cell source for cellreplacement therapies and for drug screening. Embryoid body (EB) formation from ES cells is a common method for producing different cell lineages for further applications. However, conventional techniques such as hanging drop or static suspension culture are either inherently incapable of large scale production or exhibit limited control over cell aggregation during EB formation and subsequent EB aggregation. For standardized mass EB production, a well defined scale-up platform is necessary. Recently, novel scenario methods of EB formation in hydrodynamic conditions created by bioreactor culture systems using stirred suspension systems (spinner flasks), rotating cell culture system and rotary orbital culture have allowed largescale EB formation. Their use allows for continuous monitoring and control of the physical and chemical environment which is difficult to achieve by traditional methods. This review summarizes the current state of production of EBs derived from pluripotent cells in various culture systems. Furthermore, an overview of high quality EB formation strategies coupled with systems for in vitro differentiation into various cell types to be applied in cell replacement therapy is provided in this review. Recently, new insights in induced pluripotent stem (iPS) cell technology showed that differentiation and lineage commitment are not irreversible processes and this has opened new avenues in stem cell research. These cells are equivalent to ES cells in terms of both selfrenewal and differentiation capacity. Hence, culture systems for expansion and differentiation of iPS cells can also apply methodologies developed with ES cells, although direct evidence of their use for iPS cells is still limited.
Cellular reprogramming, 2012
Mouse embryonic stem cells (ESCs) and induced pluripotent stem (iPS) cells can be used as models ... more Mouse embryonic stem cells (ESCs) and induced pluripotent stem (iPS) cells can be used as models of neuronal differentiation for the investigation of mammalian neurogenesis, pharmacological testing, and development of cell-based therapies. Recently, mouse iPS cell lines have been generated by Sleeping Beauty (SB) transposon-mediated transgenesis (SB-iPS). In this study, we determined for the first time the differentiation potential of mouse SB-iPS cells to form neuronal progenitor cells (NPCs) and neurons. Undifferentiated SB-iPS and ES cells were aggregated into embryoid bodies (EBs) and cultured in neuronal differentiation medium supplemented with 5 μM all-trans retinoic acid. Thereafter, EBs were dissociated and plated to observe further neuronal differentiation. Samples were fixed on days 10 and 14 for immunocytochemistry staining using the NPC markers Pax6 and Nestin and the neuron marker βIII-tubulin/Tuj1. Nestin-labeled cells were analyzed further by flow cytometry. Our resul...
Journal of the Medical Association of Thailand = Chotmaihet thangphaet, 2008
To determine the effect of leukemia inhibitory factor (LIF) on the quality of in vitro produced m... more To determine the effect of leukemia inhibitory factor (LIF) on the quality of in vitro produced mouse blastocyst and the efficiency of embryonic stem (ES) cell derivation. Experimental study Reproductive Medicine Unit, Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University In vivo fertilized zygotes were collected and subjected to in vitro culture in potassium simplex optimized medium (KSOM) containing 1,000 unit/ml LIF. The developmental ability of the zygote to blastocyst-stage and the cell numbers in blastocysts were evaluated Expanded blastocysts developed in different culture media were subsequently subjected to ES cell derivation. MAIN OUTCOME MEASURE (s): The influence of LIF on the quality of and the total cell numbers of blastocyst developed in vitro. Supplementation of LIF in KSOM increased the rate of hatching blastocysts (63.8% vs. 53.7%; p < 0.05) and total cell numbers (91.4 +/- 15.0 vs. 85.1 +/- 7.7; p < 0.05) compared to KSOM alo...
ABSTRACT Regenerative cell therapy against cardiovascular disease would require mass production a... more ABSTRACT Regenerative cell therapy against cardiovascular disease would require mass production and purification of specific cell types before transplantation. To enable large-scale production of embryonic stem (ES)-derived pure cardiomyocytes, we developed an animal model for a single-step scalable bioprocess that allows direct embryoid body (EB) formation in a fully controlled slow-turning lateral vessel (STLV, Synthecon, Inc., Houston, TX, USA) bioreactor following inoculation with a single cell suspension of mouse ES cells. To enhance the yield of cardiac progenitor cells, mouse ES cells (HM1; 129Sv/Ola, Magin et al. 1992 Nucl. Acids Res. 20, 3795-3796) were targeted with the cardiac-specific mouse Nkx2.5 promoter driven enhanced fluorescent green protein (EGFP). Among 15 targeted colonies, which were characterised based on morphology, the ability to form EB, EGFP expression, and in vitro differentiation ability toward cardiomyocytes, 3 lines were further evaluated for the efficiency of cardiomyocyte production. The 3 lines were cultured in STLV bioreactor and compared with classical hanging drop (HD) and static suspension culture methods. Embryonic bodies at day 3 to 8 were collected and analysed by using fluorescence-activated cell sorting for markers of pluripotency (e.g. Oct-4, SSEA1, Nanog) and cardiac (e.g. Nkx2.5, Troponin T) lineage commitments. Data was analysed by one-way ANOVA and t-tests. The results showed that both level and kinetics of Nkx2.5 expression was dependent on culture conditions. The STLV and static suspension culture methods produced higher rates of Nkx2.5-positive cells on day 5 than that of HD (70 and 54 v. 30%, respectively). The STLV method produced a highly uniform population of efficiently differentiating EB in large quantities and resulted in the highest, 10(8) yield of cardiomyocytes in a single 110-mL STLV on day 4. In conclusion, the STLV method provides a technological platform for controlled large-scale generation of ES-cell-derived cardiomyocytes for clinical and industrial applications. In vivo transplantation tests of cardiomyocytes produced via STLV are currently underway.
The aims of this study were to investigate the meiotic competence of immature porcine oocytes fol... more The aims of this study were to investigate the meiotic competence of immature porcine oocytes following open pulled straw (OPS) vitrification and warming and also to test the efficiency of the pretreatment of immature oocytes with CB prior to OPS vitrification. Experiment 1, cumulus oocyte complexes (COCs) were randomly assigned to OPS vitrification, cryoprotectant (CPA) toxicity test and non-CPA/non-vitrified controls. Experiment 2, the COCs were treated with 7.5 μg/ml CB for 30 min. The oocytes were either immediately fixed or further incubated in a CB-free medium for 5 h, non-CB treated COCs served as controls. Experiment 3, CB-pretreated COCs were vitrified. The COCs exposed to CB and vitrification solutions and non-CPA, non-CB treated oocytes served as controls. In all cases, all the COCs were matured in vitro for 40-44 h. After IVM, the oocytes were examined for the stages of nuclear maturation. Vitrifying and warming significantly reduced the capability of the immature porcine oocytes to resume and reach metaphase (M II) (21.7%), compared unfavorably with the control . This, however, was not the result from the cryoprotactants per se, since the number of CPA-treated oocytes reaching MII stage did not significantly differ from the control (76% vs. 81.8%, p>0.05). While cytochalasin B depolymerized actin cytoskeleton, this effect was completely reversible during 5 h of culture. Pretreatment with CB before the vitrification of immature porcine oocytes significantly yielded greater MII rates compared with non-CB oocytes (34.9% vs. 21.7%). In conclusion, porcine immature oocytes can be cryopreserved by OPS vitrification but it adversely affects the meiotic competence. Cytochalasin B reversibly depolymerizes actin microfilaments and improves the cryopreservability of porcine immature oocytes.
Cellular Reprogramming, 2013
Embryonic stem cells (ESCs) have the ability to form aggregates, which are called embryoid bodies... more Embryonic stem cells (ESCs) have the ability to form aggregates, which are called embryoid bodies (EBs). EBs mimic early embryonic development and are commonly produced for cardiomyogenesis. Here, we describe a method of EB formation in hydrodynamic conditions using a slow-turning lateral vessel (STLV) bioreactor and the subsequent differentiation of EBs into cardiomyocytes. EBs formed in the STLV were compared with conventional techniques, such as hanging drop (HD) or static suspension cell culture (SSC), for homogeneity of EB size, shape, proliferation, apoptosis, and in vitro cardiac differentiation. After 3 days of culture, a four-fold improvement in the yield of EB formation/mL, a six-fold enhancement in total yield of EB/mL, and a nearly 10fold reduction of cells that failed to incorporate into EBs were achieved in STLV versus SSC. During cardiac differentiation, a 1.5-to 4.2-fold increase in the area of cardiac troponin T (cTnT) per single EB in STLV versus SSC and HD was achieved. These results demonstrate that the STLV method improves the quality and quantity of ES cells to form EBs and enhances the efficiency of cardiac differentiation. We have demonstrated that the mechanical method of cell differentiation creates different microenvironments for the cells and thus influences their lineage commitments, even when genetic origin and the culture medium are the same. Ascorbic acid (ASC) improved further cardiac commitment in differentiation assays. Hence, this culture system is suitable for the production of large numbers of cells for clinical cell replacement therapies and industrial drug testing applications.
Neuroscience Letters, 2014
• Inhibition of TGF-1/ALK pathway efficiently decrease contamination of pluripotent cells in ESC... more • Inhibition of TGF-1/ALK pathway efficiently decrease contamination of pluripotent cells in ESC-derived neuronal population.
World Journal of Stem Cells, 2009
Embryonic stem (ES) cells have the ability to differ entiate into all germ layers, holding great ... more Embryonic stem (ES) cells have the ability to differ entiate into all germ layers, holding great promise not only for a model of early embryonic development but also for a robust cell source for cellreplacement therapies and for drug screening. Embryoid body (EB) formation from ES cells is a common method for producing different cell lineages for further applications. However, conventional techniques such as hanging drop or static suspension culture are either inherently incapable of large scale production or exhibit limited control over cell aggregation during EB formation and subsequent EB aggregation. For standardized mass EB production, a well defined scale-up platform is necessary. Recently, novel scenario methods of EB formation in hydrodynamic conditions created by bioreactor culture systems using stirred suspension systems (spinner flasks), rotating cell culture system and rotary orbital culture have allowed largescale EB formation. Their use allows for continuous monitoring and control of the physical and chemical environment which is difficult to achieve by traditional methods. This review summarizes the current state of production of EBs derived from pluripotent cells in various culture systems. Furthermore, an overview of high quality EB formation strategies coupled with systems for in vitro differentiation into various cell types to be applied in cell replacement therapy is provided in this review. Recently, new insights in induced pluripotent stem (iPS) cell technology showed that differentiation and lineage commitment are not irreversible processes and this has opened new avenues in stem cell research. These cells are equivalent to ES cells in terms of both selfrenewal and differentiation capacity. Hence, culture systems for expansion and differentiation of iPS cells can also apply methodologies developed with ES cells, although direct evidence of their use for iPS cells is still limited.
Molecular Biotechnology, 2014
Embryonic stem cell (ESC)-derived cardiomyocytes are a promising cell source for the screening fo... more Embryonic stem cell (ESC)-derived cardiomyocytes are a promising cell source for the screening for potential cytoprotective molecules against ischemia/reperfusion injury, however, little is known on their behavior in hypoxia/reoxygenation conditions. Here we tested the cytoprotective effect of the NO-donor SNAP and its downstream cellular pathway. Mouse ESC-derived cardiomyocytes were subjected to 150-min simulated ischemia (SI) followed by 120-min reoxygenation or corresponding non-ischemic conditions. The following treatments were applied during SI or normoxia: the NO-donor S-Nitroso-N-acetyl-D,L-penicillamine (SNAP), the protein kinase G (PKG) inhibitor, the K ATP channel blocker glibenclamide, the particulate guanylate cyclase activator brain type natriuretic peptide (BNP), and a non-specific NO synthase inhibitor (N-Nitro-L-arginine, L-NNA) alone or in different combinations. Viability of cells was assayed by propidium iodide staining. SNAP attenuated SI-induced cell death in a concentration-dependent manner, and this protection was attenuated by inhibition of either PKG or K ATP channels. However, SI-induced cell death was not A. Görbe and Z. V. Varga have contributed equally to this study.
The Thai veterinary medicine
Derivation of embryonic stem (ES) cells from parthenogenetic embryos represents a possible altern... more Derivation of embryonic stem (ES) cells from parthenogenetic embryos represents a possible alternative approach to create histocompatible cells for regenerative medicine. The objectives of this study were to establish mouse parthenogenetic ES (pES) cell lines from parthenogenetically-derived blastocysts as a model system for human and animal research and to examine pluripotency differences among the pES cell lines. We are able to report the successful establishment of four pluripotent pES cell lines from blastocysts of parthenogenetic origin (22% efficiency of pES cell line establishment). Four pES cell lines (pES#1-4) exhibited a typical ES cell morphology and expression of key pluripotency markers (ALP, Oct4, Nanog and SSEA-1). Three of the four pES cell lines have shown a high percentage of normal karyotype during long-term culture. Variability in the in vitro differentiation potential into cell types of the 3 germ layers was observed among the different pES cell lines. Three of ...