Zack Wang - Academia.edu (original) (raw)
Papers by Zack Wang
Circulation Research, 2020
Introduction: Induced pluripotent stem cells (iPSC) have a significant capacity for self-renewal ... more Introduction: Induced pluripotent stem cells (iPSC) have a significant capacity for self-renewal and can differentiate into any somatic cell type, such as megakaryocytes (MK) and vascular endothelial cells (EC). We have previously shown that iPSC-derived MKs (IPSC-MK) upregulate the gene expression levels of irreversible glycolytic enzymes committing glucose to glycolysis in comparison to their parental iPSC lines (Vaidya et al., 2016). This pattern differed from past comparisons between somatic source cells and iPSCs (Chung et al., 2007; Varum et al., 2011; Zhang et al., 2011). To further corroborate our data, we extended our analysis to iPSC-derived ECs (iPSC-EC). Methods: We studied expression of glycolytic enzyme genes in 30 human iPSC lines and paired iPSC-EC and iPSC-MK lines using RNA-seq. iPSC-EC to iPSC expression ratios for 11 glycolytic enzymes were estimated using a multilevel mixed-effect model regression, and a significance threshold of p < 0.05/11 = 0.0045 was used...
Zhonghua yi xue za zhi, 2008
OBJECTIVE To establish a sprouting embryoid body model mimicking early embryonic vasculogenesis i... more OBJECTIVE To establish a sprouting embryoid body model mimicking early embryonic vasculogenesis in human embryo. METHODS Human embryonic stem were (hESCs) were cultured on the mouse embryo fibroblasts and then were induced to differentiate to form three-dimensional EB. The hEBs were cultured in media containing various angiogenesis-related factors: vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), endostatin, angiostatin, and platelet factor (PF)-4 of different concentrations for 3 days to observe the sprouting of the hEBs. 3, 3, 3', 3'-tetramethylindo-carbocyanine perchlorate labeled acetylated low density lipoprotein (Dil-AcLDL) was added onto the hEBs foe 4 h Immunofluorescence assay was used to observe if Dil-AcLDL was absorbed and if CD31 was expressed so as to determine the existence of embryonic endothelial cells in the sprouting structures. The ideal culturing condition was analyzed. RESULTS The differentiated EBs formed sprouting structures ...
PLoS ONE, 2021
The biliary system is routinely accessed for clinical purposes via endoscopic retrograde cholangi... more The biliary system is routinely accessed for clinical purposes via endoscopic retrograde cholangiopancreatography (ERCP). We previously pioneered ERCP-mediated hydrodynamic injection in large animal models as an innovative gene delivery approach for monogenic liver diseases. However, the procedure poses potential safety concerns related mainly to liver or biliary tree injury. Here, we sought to further define biliary hydrodynamic injection parameters that are well-tolerated in a human-sized animal model. ERCP was performed in pigs, and hydrodynamic injection carried out using a novel protocol to reduce duct wall stress. Each pig was subjected to multiple repeated injections to expedite testing and judge tolerability. Different injection parameters (volume, flow rate) and injection port diameters were tested. Vital signs were monitored throughout the procedure, and liver enzyme panels were collected pre- and post-procedure. Pigs tolerated repeated biliary hydrodynamic injections with...
Blood, 2020
Human pluripotent stem cells (hPSCs) provide a powerful platform for generating functional hemato... more Human pluripotent stem cells (hPSCs) provide a powerful platform for generating functional hematopoietic cells for blood disease modeling and therapeutic testing. However, the quantity and quality of hPSC-derived blood cells remain to be improved. Here, by performing extensive single-cell transcriptomic analyses to map fate choices and gene expression programs during hematopoietic differentiation of hPSCs, we construct the first hematopoietic landscape of hPSCs at the single-cell level and identify strategies to promote hematopoietic progenitor (HP) generation from hPSCs with functional T cell potential. By focusing specifically on cell populations and molecular events involved in endothelial-to-hematopoietic transition (EHT), we compared the difference of early hematopoiesis between hPSCs and human embryos (Yang Zeng et al. Cell Research. 2019) and found aerobic metabolism was dysregulated during in-vitro-directed differentiation. The decreased oxygen metabolism program was further...
The Journal of Infectious Diseases, 1993
F1000Research, 2011
Our Machine Learning Methodology is Specifically Designed to Study Mammalian Systems Biology Abst... more Our Machine Learning Methodology is Specifically Designed to Study Mammalian Systems Biology Abstract Our Machine Learning Methodology is Specifically Designed to Study Mammalian Systems Biology Stem cells have long held tremendous promise as a versatile tool for Stem cells have long held tremendous promise as a versatile tool for biomedical research and disease therapy due to their ability to both Our cell-type specific approach is designed to control the inherent complexity of biomedical research and disease therapy due to their ability to both self renew and differentiate into distinct cell lineages The molecular mammalian systems biology by focusing on a single cell system during a specific self-renew and differentiate into distinct cell lineages. The molecular i t ti th t d i t ll lf l h l ti ll
Journal of Clinical Microbiology, 2021
Serologic point-of-care tests to detect antibodies against severe acute respiratory syndrome coro... more Serologic point-of-care tests to detect antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are an important tool in the COVID-19 pandemic. The majority of current point-of-care antibody tests developed for SARS-CoV-2 rely on lateral flow assays, but these do not offer quantitative information.
Science Advances, 2021
Hypoxia enhances hematopoiesis with functional T cell potential from human pluripotent stem cells... more Hypoxia enhances hematopoiesis with functional T cell potential from human pluripotent stem cells via arterial priming.
Cell Proliferation, 2021
This is an open access article under the terms of the Creative Commons Attribution License, which... more This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Blood, 2020
Genome-wide association studies have identified common variants associated with platelet-related ... more Genome-wide association studies have identified common variants associated with platelet-related phenotypes, but because these variants are largely intronic or intergenic, their link to platelet biology is unclear. In 290 normal subjects from the GeneSTAR Research Study (110 African Americans [AAs] and 180 European Americans [EAs]), we generated whole-genome sequence data from whole blood and RNA sequence data from extracted nonribosomal RNA from 185 induced pluripotent stem cell-derived megakaryocyte (MK) cell lines (platelet precursor cells) and 290 blood platelet samples from these subjects. Using eigenMT software to select the peak single-nucleotide polymorphism (SNP) for each expressed gene, and meta-analyzing the results of AAs and EAs, we identify (q-value < 0.05) 946 cis-expression quantitative trait loci (eQTLs) in derived MKs and 1830 cis-eQTLs in blood platelets. Among the 57 eQTLs shared between the 2 tissues, the estimated directions of effect are very consistent (98...
The COVID-19 pandemic has brought the world to a halt, with cases observed around the globe causi... more The COVID-19 pandemic has brought the world to a halt, with cases observed around the globe causing significant mortality. There is an urgent need for serological tests to detect antibodies against SARS-CoV-2, which could be used to assess the prevalence of infection, as well as ascertain individuals who may be protected from future infection. Current serological tests developed for SARS-CoV-2 rely on traditional technologies such as enzyme-linked immunosorbent assays (ELISA) and lateral flow assays, which may lack scalability to meet the demand of hundreds of millions of antibody tests in the coming year. Herein, we present an alternative method of antibody testing that just depends on one protein reagent being added to patient serum/plasma or whole blood and a short five-minute assay time. A novel fusion protein was designed that binds red blood cells (RBC) via a single-chain variable fragment (scFv) against the H antigen and displays the receptor-binding domain (RBD) of SARS-CoV-...
Experimental Hematology, 2018
Myelodysplastic syndrome (MDS) is a disease caused by an ineffective hematopoiesis. MDS can devel... more Myelodysplastic syndrome (MDS) is a disease caused by an ineffective hematopoiesis. MDS can develop into acute myeloid leukemia (AML), an aggressive blood cancer of the myeloid lineage. AML patients have a poor prognosis despite treatment with intensive chemotherapy, hence alternative therapies are needed. Gfi1 (Growth factor independence 1) is a transcriptional repressor which, among other functions, recruits HDAC1 and 2 (histone deacetylase 1 and 2) to its target genes. Low Gfi1 levels (Gfi1-KD) in MDS and AML blasts are associated with poor patient survival and AML development in murine models of human AML. To understand how reduced Gfi1 levels contribute to AML, we analyzed the effect of Gfi1-KD on normal and malignant hematopoietic stem cells (HSCs) in mice. Gfi1-KD led to increased self-renewal in HSCs due to increased symmetric division. In addition, Gfi1-KD resulted in the numeric expansion of leukemic stem cells (LSCs). On a molecular level, expression of Gfi1 leads to the removal of acetyl groups at H3K9. Gfi1-KD therefore resulted in increased H3K9 acetylation at its target genes, causing increased target gene expression. A number of these target genes were oncogenes, explaining why Gfi1-KD promotes AML. We thus hypothesized that the administration of HAT (histone acetyltransferase) inhibitors could be beneficial for MDS/AML patients with reduced Gfi1 expression as it could reverse the increased acetylation of H3K9. Curcumin is a HAT inhibitor which is used as a spice and has so far no known toxic side effects. To study the effect of Curcumin on MDS/AML development, we crossed the well-established murine model of human MDS/AML, NUP98-HOXD13, with GFI1-WT or GFI1-KD mice. The different groups were treated with either curcumin or as a control with 5-Azacytidine (Aza), a DNA methyltransferase inhibitor used in the clinic for the treatment of MDS patients. Curcumin effectively prevented the development of AML in mice with low Gfi1 expression, while the control did not. Overall, our data suggest that Gfi1 regulates the epigenetic state of HSCs and LSCs and thus their stemness in a dose-dependent manner and that Curcumin could be used as a targeted therapy.
Blood, 2005
Human embryonic stem cell (hESC) lines provide not only an unique system for studying human embry... more Human embryonic stem cell (hESC) lines provide not only an unique system for studying human embryonic development, but also have enormous potential as a source of therapeutic tissues, including those of vascular and hematopoietic lineages. However, it is difficult to generate large-scale of relatively pure population of specific lineage cells derived from hESCs. Here, we describe a 2-dimensional (2-D) culture system for hESC differentiation to generate hematopoietic and endothelial cells in vitro without embryoid body (EB) formation (3-D). To initial differentiation, hESCs (H1 and H9 cell lines) were placed in serum-containing media for 9–10 days without supplement of additional growth factors. Gene expression analysis of 2-D differentiation indicated that Oct-4, a pluripotent gene, was decreased, while genes for GATA-2 and CD31, were increased, suggesting that differentiated hESCs contain hematopoietic and endothelial progenitors. Comparing with 3-D EB formation, 2-D differentiatio...
Journal of Cellular Physiology, 2020
Ischemic heart disease and congestive heart failure are major contributors to high morbidity and ... more Ischemic heart disease and congestive heart failure are major contributors to high morbidity and mortality. Approximately 1.5 million cases of myocardial infarction (MI) occur annually in the United States; the yearly incidence rate is approximately 600 cases per 100,000 people. Although significant progress to improve survival rate has been made by medications and implantable medical devices, damaged cardiomyocytes are unable to be recovered by current treatment strategies. After almost two decades of research, stem cell therapy has become a very promising approach to generate new cardiomyocytes and enhance function of heart. Along with clinical trials with stem cells conducted in cardiac regeneration, concerns regarding safety and potential risks have emerged. One of the contentious issues is electrical dysfunctions of cardiomyocytes and cardiac arrhythmia after stem cell therapy. In this review, we focus on the cell sources currently used for stem cell therapy and discuss related arrhythmogenic risk.
Blood, 2004
Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) is widely used as endothelial marker... more Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) is widely used as endothelial marker during vasculogenesis and angiogenesis from embryonic stem (ES) cells. Here, we demonstrate that undifferentiated ES cells expressed high level PECAM-1, but low levels of other endothelium markers, including VE-cadherin, Flk-1 and CD34. Kinetic analysis revealed that the expression of PECAM-1 was sharply down-regulated during early ES cell differentiation. PECAM-1 was expressed in the vascular-like area along with other endothelium makers during differentiation of ES cells. In the absence of angiogenic growth factors, embryonic bodies (EBs) were lack of vascular-like structures, and PECAM-1 was detected in the cells without organized structure. Furthermore, undifferentiated ES cells were found to express all eight known alternatively spliced isoforms of PECAM-1, among them, the expression of PECAM-1 isoforms lacking exon 15, (D15), or 14&15 (D14&15) was predominant. Only the expression ...
Blood, 2009
3037 Poster Board II-1013 The generation of vascular cells from pluripotent stem cells, including... more 3037 Poster Board II-1013 The generation of vascular cells from pluripotent stem cells, including human embryonic stem cells (hESCs) and induced pluripotent stem (iPS) cells, may facilitate tissue transplantation, reperfusion of ischemic tissues, and treatment of pathologies in which endothelial cell dysfunction exists. The signals that direct pluripotent stem cell differentiation into lineage-specific cells remain largely unknown. To identify vascular progenitor cells during hESC differentiation, we characterized various subpopulations that may differentiate into endothelial cells (ECs) and smooth muscle cells (SMCs). In our newly established serum-free medium, hESCs sequentially differentiated into CD34+CD31-, CD34+CD31+, and then CD34-CD31+ cells. Real-time PCR analysis indicated that EC- and SMC-specific genes, including VEGFR2, Tie2, VE-Cad, vWF, SMA, SM22-alpha, calponin, and caldesmon, were expressed significantly high in CD34+CD31+ cells. Furthermore, the CD34+CD31+ cells th...
Blood, 2010
2579 Slug belongs to the highly conserved Slug/Snail family of zinc-finger transcription factors ... more 2579 Slug belongs to the highly conserved Slug/Snail family of zinc-finger transcription factors found in diverse species ranging from C. elegans to humans. Our previous study has already demonstrated that Slug serves intrinsic role in the regulation of HSC fates (Blood, 2010, 115(9):1709-1717). There is bone defect in Slug-/- mice, while Slug is expressed in osteoblast cells which are niche cells for HSCs, suggesting that Slug might be also extrinsically required for HSC regulation. To address this issue, the following experiments and assays were performed. Firstly, the cell proliferation of wild-type (WT) hematopoietic stem/progenitor cells (HS/PCs) (loaded with carboxyfluorescein succinimidyl ester (CFSE)) placed on bone marrow (BM) stromal layers (prepared from either Slug-/- or WT mice) was analyzed. Co-culture proliferation assay illustrated a clear increasing in the ability of Slug-/-BM stroma to support normal proliferation of WT HS/PCs, suggesting a significant and qualitat...
Circulation Research, 2020
Introduction: Induced pluripotent stem cells (iPSC) have a significant capacity for self-renewal ... more Introduction: Induced pluripotent stem cells (iPSC) have a significant capacity for self-renewal and can differentiate into any somatic cell type, such as megakaryocytes (MK) and vascular endothelial cells (EC). We have previously shown that iPSC-derived MKs (IPSC-MK) upregulate the gene expression levels of irreversible glycolytic enzymes committing glucose to glycolysis in comparison to their parental iPSC lines (Vaidya et al., 2016). This pattern differed from past comparisons between somatic source cells and iPSCs (Chung et al., 2007; Varum et al., 2011; Zhang et al., 2011). To further corroborate our data, we extended our analysis to iPSC-derived ECs (iPSC-EC). Methods: We studied expression of glycolytic enzyme genes in 30 human iPSC lines and paired iPSC-EC and iPSC-MK lines using RNA-seq. iPSC-EC to iPSC expression ratios for 11 glycolytic enzymes were estimated using a multilevel mixed-effect model regression, and a significance threshold of p < 0.05/11 = 0.0045 was used...
Zhonghua yi xue za zhi, 2008
OBJECTIVE To establish a sprouting embryoid body model mimicking early embryonic vasculogenesis i... more OBJECTIVE To establish a sprouting embryoid body model mimicking early embryonic vasculogenesis in human embryo. METHODS Human embryonic stem were (hESCs) were cultured on the mouse embryo fibroblasts and then were induced to differentiate to form three-dimensional EB. The hEBs were cultured in media containing various angiogenesis-related factors: vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), endostatin, angiostatin, and platelet factor (PF)-4 of different concentrations for 3 days to observe the sprouting of the hEBs. 3, 3, 3', 3'-tetramethylindo-carbocyanine perchlorate labeled acetylated low density lipoprotein (Dil-AcLDL) was added onto the hEBs foe 4 h Immunofluorescence assay was used to observe if Dil-AcLDL was absorbed and if CD31 was expressed so as to determine the existence of embryonic endothelial cells in the sprouting structures. The ideal culturing condition was analyzed. RESULTS The differentiated EBs formed sprouting structures ...
PLoS ONE, 2021
The biliary system is routinely accessed for clinical purposes via endoscopic retrograde cholangi... more The biliary system is routinely accessed for clinical purposes via endoscopic retrograde cholangiopancreatography (ERCP). We previously pioneered ERCP-mediated hydrodynamic injection in large animal models as an innovative gene delivery approach for monogenic liver diseases. However, the procedure poses potential safety concerns related mainly to liver or biliary tree injury. Here, we sought to further define biliary hydrodynamic injection parameters that are well-tolerated in a human-sized animal model. ERCP was performed in pigs, and hydrodynamic injection carried out using a novel protocol to reduce duct wall stress. Each pig was subjected to multiple repeated injections to expedite testing and judge tolerability. Different injection parameters (volume, flow rate) and injection port diameters were tested. Vital signs were monitored throughout the procedure, and liver enzyme panels were collected pre- and post-procedure. Pigs tolerated repeated biliary hydrodynamic injections with...
Blood, 2020
Human pluripotent stem cells (hPSCs) provide a powerful platform for generating functional hemato... more Human pluripotent stem cells (hPSCs) provide a powerful platform for generating functional hematopoietic cells for blood disease modeling and therapeutic testing. However, the quantity and quality of hPSC-derived blood cells remain to be improved. Here, by performing extensive single-cell transcriptomic analyses to map fate choices and gene expression programs during hematopoietic differentiation of hPSCs, we construct the first hematopoietic landscape of hPSCs at the single-cell level and identify strategies to promote hematopoietic progenitor (HP) generation from hPSCs with functional T cell potential. By focusing specifically on cell populations and molecular events involved in endothelial-to-hematopoietic transition (EHT), we compared the difference of early hematopoiesis between hPSCs and human embryos (Yang Zeng et al. Cell Research. 2019) and found aerobic metabolism was dysregulated during in-vitro-directed differentiation. The decreased oxygen metabolism program was further...
The Journal of Infectious Diseases, 1993
F1000Research, 2011
Our Machine Learning Methodology is Specifically Designed to Study Mammalian Systems Biology Abst... more Our Machine Learning Methodology is Specifically Designed to Study Mammalian Systems Biology Abstract Our Machine Learning Methodology is Specifically Designed to Study Mammalian Systems Biology Stem cells have long held tremendous promise as a versatile tool for Stem cells have long held tremendous promise as a versatile tool for biomedical research and disease therapy due to their ability to both Our cell-type specific approach is designed to control the inherent complexity of biomedical research and disease therapy due to their ability to both self renew and differentiate into distinct cell lineages The molecular mammalian systems biology by focusing on a single cell system during a specific self-renew and differentiate into distinct cell lineages. The molecular i t ti th t d i t ll lf l h l ti ll
Journal of Clinical Microbiology, 2021
Serologic point-of-care tests to detect antibodies against severe acute respiratory syndrome coro... more Serologic point-of-care tests to detect antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are an important tool in the COVID-19 pandemic. The majority of current point-of-care antibody tests developed for SARS-CoV-2 rely on lateral flow assays, but these do not offer quantitative information.
Science Advances, 2021
Hypoxia enhances hematopoiesis with functional T cell potential from human pluripotent stem cells... more Hypoxia enhances hematopoiesis with functional T cell potential from human pluripotent stem cells via arterial priming.
Cell Proliferation, 2021
This is an open access article under the terms of the Creative Commons Attribution License, which... more This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Blood, 2020
Genome-wide association studies have identified common variants associated with platelet-related ... more Genome-wide association studies have identified common variants associated with platelet-related phenotypes, but because these variants are largely intronic or intergenic, their link to platelet biology is unclear. In 290 normal subjects from the GeneSTAR Research Study (110 African Americans [AAs] and 180 European Americans [EAs]), we generated whole-genome sequence data from whole blood and RNA sequence data from extracted nonribosomal RNA from 185 induced pluripotent stem cell-derived megakaryocyte (MK) cell lines (platelet precursor cells) and 290 blood platelet samples from these subjects. Using eigenMT software to select the peak single-nucleotide polymorphism (SNP) for each expressed gene, and meta-analyzing the results of AAs and EAs, we identify (q-value < 0.05) 946 cis-expression quantitative trait loci (eQTLs) in derived MKs and 1830 cis-eQTLs in blood platelets. Among the 57 eQTLs shared between the 2 tissues, the estimated directions of effect are very consistent (98...
The COVID-19 pandemic has brought the world to a halt, with cases observed around the globe causi... more The COVID-19 pandemic has brought the world to a halt, with cases observed around the globe causing significant mortality. There is an urgent need for serological tests to detect antibodies against SARS-CoV-2, which could be used to assess the prevalence of infection, as well as ascertain individuals who may be protected from future infection. Current serological tests developed for SARS-CoV-2 rely on traditional technologies such as enzyme-linked immunosorbent assays (ELISA) and lateral flow assays, which may lack scalability to meet the demand of hundreds of millions of antibody tests in the coming year. Herein, we present an alternative method of antibody testing that just depends on one protein reagent being added to patient serum/plasma or whole blood and a short five-minute assay time. A novel fusion protein was designed that binds red blood cells (RBC) via a single-chain variable fragment (scFv) against the H antigen and displays the receptor-binding domain (RBD) of SARS-CoV-...
Experimental Hematology, 2018
Myelodysplastic syndrome (MDS) is a disease caused by an ineffective hematopoiesis. MDS can devel... more Myelodysplastic syndrome (MDS) is a disease caused by an ineffective hematopoiesis. MDS can develop into acute myeloid leukemia (AML), an aggressive blood cancer of the myeloid lineage. AML patients have a poor prognosis despite treatment with intensive chemotherapy, hence alternative therapies are needed. Gfi1 (Growth factor independence 1) is a transcriptional repressor which, among other functions, recruits HDAC1 and 2 (histone deacetylase 1 and 2) to its target genes. Low Gfi1 levels (Gfi1-KD) in MDS and AML blasts are associated with poor patient survival and AML development in murine models of human AML. To understand how reduced Gfi1 levels contribute to AML, we analyzed the effect of Gfi1-KD on normal and malignant hematopoietic stem cells (HSCs) in mice. Gfi1-KD led to increased self-renewal in HSCs due to increased symmetric division. In addition, Gfi1-KD resulted in the numeric expansion of leukemic stem cells (LSCs). On a molecular level, expression of Gfi1 leads to the removal of acetyl groups at H3K9. Gfi1-KD therefore resulted in increased H3K9 acetylation at its target genes, causing increased target gene expression. A number of these target genes were oncogenes, explaining why Gfi1-KD promotes AML. We thus hypothesized that the administration of HAT (histone acetyltransferase) inhibitors could be beneficial for MDS/AML patients with reduced Gfi1 expression as it could reverse the increased acetylation of H3K9. Curcumin is a HAT inhibitor which is used as a spice and has so far no known toxic side effects. To study the effect of Curcumin on MDS/AML development, we crossed the well-established murine model of human MDS/AML, NUP98-HOXD13, with GFI1-WT or GFI1-KD mice. The different groups were treated with either curcumin or as a control with 5-Azacytidine (Aza), a DNA methyltransferase inhibitor used in the clinic for the treatment of MDS patients. Curcumin effectively prevented the development of AML in mice with low Gfi1 expression, while the control did not. Overall, our data suggest that Gfi1 regulates the epigenetic state of HSCs and LSCs and thus their stemness in a dose-dependent manner and that Curcumin could be used as a targeted therapy.
Blood, 2005
Human embryonic stem cell (hESC) lines provide not only an unique system for studying human embry... more Human embryonic stem cell (hESC) lines provide not only an unique system for studying human embryonic development, but also have enormous potential as a source of therapeutic tissues, including those of vascular and hematopoietic lineages. However, it is difficult to generate large-scale of relatively pure population of specific lineage cells derived from hESCs. Here, we describe a 2-dimensional (2-D) culture system for hESC differentiation to generate hematopoietic and endothelial cells in vitro without embryoid body (EB) formation (3-D). To initial differentiation, hESCs (H1 and H9 cell lines) were placed in serum-containing media for 9–10 days without supplement of additional growth factors. Gene expression analysis of 2-D differentiation indicated that Oct-4, a pluripotent gene, was decreased, while genes for GATA-2 and CD31, were increased, suggesting that differentiated hESCs contain hematopoietic and endothelial progenitors. Comparing with 3-D EB formation, 2-D differentiatio...
Journal of Cellular Physiology, 2020
Ischemic heart disease and congestive heart failure are major contributors to high morbidity and ... more Ischemic heart disease and congestive heart failure are major contributors to high morbidity and mortality. Approximately 1.5 million cases of myocardial infarction (MI) occur annually in the United States; the yearly incidence rate is approximately 600 cases per 100,000 people. Although significant progress to improve survival rate has been made by medications and implantable medical devices, damaged cardiomyocytes are unable to be recovered by current treatment strategies. After almost two decades of research, stem cell therapy has become a very promising approach to generate new cardiomyocytes and enhance function of heart. Along with clinical trials with stem cells conducted in cardiac regeneration, concerns regarding safety and potential risks have emerged. One of the contentious issues is electrical dysfunctions of cardiomyocytes and cardiac arrhythmia after stem cell therapy. In this review, we focus on the cell sources currently used for stem cell therapy and discuss related arrhythmogenic risk.
Blood, 2004
Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) is widely used as endothelial marker... more Platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) is widely used as endothelial marker during vasculogenesis and angiogenesis from embryonic stem (ES) cells. Here, we demonstrate that undifferentiated ES cells expressed high level PECAM-1, but low levels of other endothelium markers, including VE-cadherin, Flk-1 and CD34. Kinetic analysis revealed that the expression of PECAM-1 was sharply down-regulated during early ES cell differentiation. PECAM-1 was expressed in the vascular-like area along with other endothelium makers during differentiation of ES cells. In the absence of angiogenic growth factors, embryonic bodies (EBs) were lack of vascular-like structures, and PECAM-1 was detected in the cells without organized structure. Furthermore, undifferentiated ES cells were found to express all eight known alternatively spliced isoforms of PECAM-1, among them, the expression of PECAM-1 isoforms lacking exon 15, (D15), or 14&15 (D14&15) was predominant. Only the expression ...
Blood, 2009
3037 Poster Board II-1013 The generation of vascular cells from pluripotent stem cells, including... more 3037 Poster Board II-1013 The generation of vascular cells from pluripotent stem cells, including human embryonic stem cells (hESCs) and induced pluripotent stem (iPS) cells, may facilitate tissue transplantation, reperfusion of ischemic tissues, and treatment of pathologies in which endothelial cell dysfunction exists. The signals that direct pluripotent stem cell differentiation into lineage-specific cells remain largely unknown. To identify vascular progenitor cells during hESC differentiation, we characterized various subpopulations that may differentiate into endothelial cells (ECs) and smooth muscle cells (SMCs). In our newly established serum-free medium, hESCs sequentially differentiated into CD34+CD31-, CD34+CD31+, and then CD34-CD31+ cells. Real-time PCR analysis indicated that EC- and SMC-specific genes, including VEGFR2, Tie2, VE-Cad, vWF, SMA, SM22-alpha, calponin, and caldesmon, were expressed significantly high in CD34+CD31+ cells. Furthermore, the CD34+CD31+ cells th...
Blood, 2010
2579 Slug belongs to the highly conserved Slug/Snail family of zinc-finger transcription factors ... more 2579 Slug belongs to the highly conserved Slug/Snail family of zinc-finger transcription factors found in diverse species ranging from C. elegans to humans. Our previous study has already demonstrated that Slug serves intrinsic role in the regulation of HSC fates (Blood, 2010, 115(9):1709-1717). There is bone defect in Slug-/- mice, while Slug is expressed in osteoblast cells which are niche cells for HSCs, suggesting that Slug might be also extrinsically required for HSC regulation. To address this issue, the following experiments and assays were performed. Firstly, the cell proliferation of wild-type (WT) hematopoietic stem/progenitor cells (HS/PCs) (loaded with carboxyfluorescein succinimidyl ester (CFSE)) placed on bone marrow (BM) stromal layers (prepared from either Slug-/- or WT mice) was analyzed. Co-culture proliferation assay illustrated a clear increasing in the ability of Slug-/-BM stroma to support normal proliferation of WT HS/PCs, suggesting a significant and qualitat...