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Ann M Simpson - One of the best experts on this subject based on the ideXlab platform.
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reversal of diabetes following transplantation of an insulin secreting human Liver Cell line melligen Cells
Molecular therapy. Methods & clinical development, 2015Co-Authors: Janet Lawandi, Anne M Swan, Dora Ling, Najah T Nassif, Fraser R. Torpy, Bronwyn A Obrien, P.f. Williams, Ann M SimpsonAbstract:As an alternative to the transplantation of islets, a human Liver Cell line has been genetically engineered to reverse type 1 diabetes (TID). The initial Liver Cell line (Huh7ins) commenced secretion of insulin in response to a glucose concentration of 2.5 mmol/l. After transfection of the Huh7ins Cells with human islet glucokinase, the resultant Melligen Cells secreted insulin in response to glucose within the physiological range; commencing at 4.25 mmol/l. Melligen Cells exhibited increased glucokinase enzymatic activity in response to physiological glucose concentrations, as compared with Huh7ins Cells. When transplanted into diabetic immunoincompetent mice, Melligen Cells restored normoglycemia. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that both Cell lines expressed a range of β-Cell transcription factors and pancreatic hormones. Exposure of Melligen and Huh7ins Cells to proinflammatory cytokines (TNF-α, IL-1β, and IFN-γ) affected neither their viability nor their ability to secrete insulin to glucose. Gene expression (microarray and qRT-PCR) analyses indicated the survival of Melligen Cells in the presence of known β-Cell cytotoxins was associated with the expression of NF-κB and antiapoptotic genes (such as BIRC3). This study describes the successful generation of an artificial β-Cell line, which, if encapsulated to avoid allograft rejection, may offer a clinically applicable cure for T1D.
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reversal of diabetes following transplantation of an insulin secreting human Liver Cell line melligen Cells
Molecular therapy. Methods & clinical development, 2015Co-Authors: Janet Lawandi, Anne M Swan, Dora Ling, Najah T Nassif, Fraser R. Torpy, Bronwyn A Obrien, P.f. Williams, Ann M SimpsonAbstract:As an alternative to the transplantation of islets, a human Liver Cell line has been genetically engineered to reverse type 1 diabetes (TID). The initial Liver Cell line (Huh7ins) commenced secretion of insulin in response to a glucose concentration of 2.5 mmol/l. After transfection of the Huh7ins Cells with human islet glucokinase, the resultant Melligen Cells secreted insulin in response to glucose within the physiological range; commencing at 4.25 mmol/l. Melligen Cells exhibited increased glucokinase enzymatic activity in response to physiological glucose concentrations, as compared with Huh7ins Cells. When transplanted into diabetic immunoincompetent mice, Melligen Cells restored normoglycemia. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that both Cell lines expressed a range of β-Cell transcription factors and pancreatic hormones. Exposure of Melligen and Huh7ins Cells to proinflammatory cytokines (TNF-α, IL-1β, and IFN-γ) affected neither their viability nor their ability to secrete insulin to glucose. Gene expression (microarray and qRT-PCR) analyses indicated the survival of Melligen Cells in the presence of known β-Cell cytotoxins was associated with the expression of NF-κB and antiapoptotic genes (such as BIRC3). This study describes the successful generation of an artificial β-Cell line, which, if encapsulated to avoid allograft rejection, may offer a clinically applicable cure for T1D.
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function of a genetically modified human Liver Cell line that stores processes and secretes insulin
Gene Therapy, 2003Co-Authors: Bernard E Tuch, Muhammad T Tabiin, Sara Holman, Anne M Swan, R K Humphrey, B Szymanska, David J Gross, Glenn M. Marshall, Ann M SimpsonAbstract:Function of a genetically modified human Liver Cell line that stores, processes and secretes insulin
Shaufeng Chang - One of the best experts on this subject based on the ideXlab platform.
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Liver Cell patterning lab chip mimicking the morphology of Liver lobule tissue
Lab on a Chip, 2013Co-Authors: Chenta Ho, Rongjhe Chen, Chungkuang Chin, Songen Gong, Hwanyou Chang, Hweiling Peng, Shaufeng ChangAbstract:A lobule-mimetic Cell-patterning technique for on-chip reconstruction of centimetre-scale Liver tissue of heterogeneous hepatic and endothelial Cells via an enhanced field-induced dielectrophoresis (DEP) trap is demonstrated and reported. By mimicking the basic morphology of Liver tissue, the classic hepatic lobule, the lobule-mimetic-stellate-electrodes array was designed for Cell patterning. Through DEP manipulation, well-defined and enhanced spatial electric field gradients were created for in-parallel manipulation of massive individual Cells. With this Liver-Cell patterning labchip design, the original randomly distributed hepatic and endothelial Cells inside the microfluidic chamber can be manipulated separately and aligned into the desired pattern that mimicks the morphology of Liver lobule tissue. Experimental results showed that both hepatic and endothelial Cells were orderly guided, snared, and aligned along the field-induced orientation to form the lobule-mimetic pattern. About 95% Cell viability of hepatic and endothelial Cells was also observed after Cell-patterning demonstration via a fluorescent assay technique. The Liver function of CYP450-1A1 enzyme activity showed an 80% enhancement for our engineered Liver tissue (HepG2+HUVECs) compared to the non-patterned pure HepG2 for two-day culturing.
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Liver Cell patterning lab chip mimicking the morphology of Liver lobule tissue
Lab on a Chip, 2013Co-Authors: Chenta Ho, Rongjhe Chen, Chungkuang Chin, Songen Gong, Hwanyou Chang, Hweiling Peng, Shaufeng ChangAbstract:A lobule-mimetic Cell-patterning technique for on-chip reconstruction of centimetre-scale Liver tissue of heterogeneous hepatic and endothelial Cells via an enhanced field-induced dielectrophoresis (DEP) trap is demonstrated and reported. By mimicking the basic morphology of Liver tissue, the classic hepatic lobule, the lobule-mimetic-stellate-electrodes array was designed for Cell patterning. Through DEP manipulation, well-defined and enhanced spatial electric field gradients were created for in-parallel manipulation of massive individual Cells. With this Liver-Cell patterning labchip design, the original randomly distributed hepatic and endothelial Cells inside the microfluidic chamber can be manipulated separately and aligned into the desired pattern that mimicks the morphology of Liver lobule tissue. Experimental results showed that both hepatic and endothelial Cells were orderly guided, snared, and aligned along the field-induced orientation to form the lobule-mimetic pattern. About 95% Cell viability of hepatic and endothelial Cells was also observed after Cell-patterning demonstration via a fluorescent assay technique. The Liver function of CYP450-1A1 enzyme activity showed an 80% enhancement for our engineered Liver tissue (HepG2+HUVECs) compared to the non-patterned pure HepG2 for two-day culturing.
Anne M Swan - One of the best experts on this subject based on the ideXlab platform.
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reversal of diabetes following transplantation of an insulin secreting human Liver Cell line melligen Cells
Molecular therapy. Methods & clinical development, 2015Co-Authors: Janet Lawandi, Anne M Swan, Dora Ling, Najah T Nassif, Fraser R. Torpy, Bronwyn A Obrien, P.f. Williams, Ann M SimpsonAbstract:As an alternative to the transplantation of islets, a human Liver Cell line has been genetically engineered to reverse type 1 diabetes (TID). The initial Liver Cell line (Huh7ins) commenced secretion of insulin in response to a glucose concentration of 2.5 mmol/l. After transfection of the Huh7ins Cells with human islet glucokinase, the resultant Melligen Cells secreted insulin in response to glucose within the physiological range; commencing at 4.25 mmol/l. Melligen Cells exhibited increased glucokinase enzymatic activity in response to physiological glucose concentrations, as compared with Huh7ins Cells. When transplanted into diabetic immunoincompetent mice, Melligen Cells restored normoglycemia. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that both Cell lines expressed a range of β-Cell transcription factors and pancreatic hormones. Exposure of Melligen and Huh7ins Cells to proinflammatory cytokines (TNF-α, IL-1β, and IFN-γ) affected neither their viability nor their ability to secrete insulin to glucose. Gene expression (microarray and qRT-PCR) analyses indicated the survival of Melligen Cells in the presence of known β-Cell cytotoxins was associated with the expression of NF-κB and antiapoptotic genes (such as BIRC3). This study describes the successful generation of an artificial β-Cell line, which, if encapsulated to avoid allograft rejection, may offer a clinically applicable cure for T1D.
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reversal of diabetes following transplantation of an insulin secreting human Liver Cell line melligen Cells
Molecular therapy. Methods & clinical development, 2015Co-Authors: Janet Lawandi, Anne M Swan, Dora Ling, Najah T Nassif, Fraser R. Torpy, Bronwyn A Obrien, P.f. Williams, Ann M SimpsonAbstract:As an alternative to the transplantation of islets, a human Liver Cell line has been genetically engineered to reverse type 1 diabetes (TID). The initial Liver Cell line (Huh7ins) commenced secretion of insulin in response to a glucose concentration of 2.5 mmol/l. After transfection of the Huh7ins Cells with human islet glucokinase, the resultant Melligen Cells secreted insulin in response to glucose within the physiological range; commencing at 4.25 mmol/l. Melligen Cells exhibited increased glucokinase enzymatic activity in response to physiological glucose concentrations, as compared with Huh7ins Cells. When transplanted into diabetic immunoincompetent mice, Melligen Cells restored normoglycemia. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that both Cell lines expressed a range of β-Cell transcription factors and pancreatic hormones. Exposure of Melligen and Huh7ins Cells to proinflammatory cytokines (TNF-α, IL-1β, and IFN-γ) affected neither their viability nor their ability to secrete insulin to glucose. Gene expression (microarray and qRT-PCR) analyses indicated the survival of Melligen Cells in the presence of known β-Cell cytotoxins was associated with the expression of NF-κB and antiapoptotic genes (such as BIRC3). This study describes the successful generation of an artificial β-Cell line, which, if encapsulated to avoid allograft rejection, may offer a clinically applicable cure for T1D.
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function of a genetically modified human Liver Cell line that stores processes and secretes insulin
Gene Therapy, 2003Co-Authors: Bernard E Tuch, Muhammad T Tabiin, Sara Holman, Anne M Swan, R K Humphrey, B Szymanska, David J Gross, Glenn M. Marshall, Ann M SimpsonAbstract:Function of a genetically modified human Liver Cell line that stores, processes and secretes insulin
Janet Lawandi - One of the best experts on this subject based on the ideXlab platform.
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reversal of diabetes following transplantation of an insulin secreting human Liver Cell line melligen Cells
Molecular therapy. Methods & clinical development, 2015Co-Authors: Janet Lawandi, Anne M Swan, Dora Ling, Najah T Nassif, Fraser R. Torpy, Bronwyn A Obrien, P.f. Williams, Ann M SimpsonAbstract:As an alternative to the transplantation of islets, a human Liver Cell line has been genetically engineered to reverse type 1 diabetes (TID). The initial Liver Cell line (Huh7ins) commenced secretion of insulin in response to a glucose concentration of 2.5 mmol/l. After transfection of the Huh7ins Cells with human islet glucokinase, the resultant Melligen Cells secreted insulin in response to glucose within the physiological range; commencing at 4.25 mmol/l. Melligen Cells exhibited increased glucokinase enzymatic activity in response to physiological glucose concentrations, as compared with Huh7ins Cells. When transplanted into diabetic immunoincompetent mice, Melligen Cells restored normoglycemia. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that both Cell lines expressed a range of β-Cell transcription factors and pancreatic hormones. Exposure of Melligen and Huh7ins Cells to proinflammatory cytokines (TNF-α, IL-1β, and IFN-γ) affected neither their viability nor their ability to secrete insulin to glucose. Gene expression (microarray and qRT-PCR) analyses indicated the survival of Melligen Cells in the presence of known β-Cell cytotoxins was associated with the expression of NF-κB and antiapoptotic genes (such as BIRC3). This study describes the successful generation of an artificial β-Cell line, which, if encapsulated to avoid allograft rejection, may offer a clinically applicable cure for T1D.
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reversal of diabetes following transplantation of an insulin secreting human Liver Cell line melligen Cells
Molecular therapy. Methods & clinical development, 2015Co-Authors: Janet Lawandi, Anne M Swan, Dora Ling, Najah T Nassif, Fraser R. Torpy, Bronwyn A Obrien, P.f. Williams, Ann M SimpsonAbstract:As an alternative to the transplantation of islets, a human Liver Cell line has been genetically engineered to reverse type 1 diabetes (TID). The initial Liver Cell line (Huh7ins) commenced secretion of insulin in response to a glucose concentration of 2.5 mmol/l. After transfection of the Huh7ins Cells with human islet glucokinase, the resultant Melligen Cells secreted insulin in response to glucose within the physiological range; commencing at 4.25 mmol/l. Melligen Cells exhibited increased glucokinase enzymatic activity in response to physiological glucose concentrations, as compared with Huh7ins Cells. When transplanted into diabetic immunoincompetent mice, Melligen Cells restored normoglycemia. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that both Cell lines expressed a range of β-Cell transcription factors and pancreatic hormones. Exposure of Melligen and Huh7ins Cells to proinflammatory cytokines (TNF-α, IL-1β, and IFN-γ) affected neither their viability nor their ability to secrete insulin to glucose. Gene expression (microarray and qRT-PCR) analyses indicated the survival of Melligen Cells in the presence of known β-Cell cytotoxins was associated with the expression of NF-κB and antiapoptotic genes (such as BIRC3). This study describes the successful generation of an artificial β-Cell line, which, if encapsulated to avoid allograft rejection, may offer a clinically applicable cure for T1D.
Ruurdtje Hoekstra - One of the best experts on this subject based on the ideXlab platform.
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a practice changing culture method relying on shaking substantially increases mitochondrial energy metabolism and functionality of human Liver Cell lines
PLOS ONE, 2018Co-Authors: Aziza A. A. Adam, Manon E. Wildenberg, Vincent A Van Der Mark, Ronald Oude P J Elferink, Robert A F M Chamuleau, Joanne M Donkers, Ruurdtje HoekstraAbstract:Practice-changing culturing techniques of hepatocytes are highly required to increase their differentiation. Previously, we found that human Liver Cell lines HepaRG and C3A acquire higher functionality and increased mitochondrial biogenesis when cultured in the AMC-Bioartificial Liver (BAL). Dynamic medium flow (DMF) is one of the major contributors to this stimulatory effect. Recently, we found that DMF-culturing by shaking of HepaRG monolayers resulted in higher mitochondrial biogenesis. Here we further investigated the effect of DMF-culturing on energy metabolism and hepatic functionality of HepaRG and C3A monolayers. HepaRG and C3A DMF-monolayers were incubated with orbital shaking at 60 rpm during the differentiation phase, while control monolayers were maintained statically. Subsequently, energy metabolism and hepatic functionality were compared between static and DMF-cultures. DMF-culturing of HepaRG Cells substantially increased hepatic differentiation; transcript levels of hepatic structural genes and hepatic transcription regulators were increased up to 15-fold (Cytochrome P450 3A4) and nuclear translocation of hepatic transcription factor CEBPα was stimulated. Accordingly, hepatic functions were positively affected, including ammonia elimination, urea production, bile acid production, and CYP3A4 activity. DMF-culturing shifted energy metabolism from aerobic glycolysis towards oxidative phosphorylation, as indicated by a decline in lactate production and glucose consumption, and an increase in oxygen consumption. Similarly, DMF-culturing increased mitochondrial energy metabolism and hepatic functionality of C3A Cells. In conclusion, simple shaking of monolayer cultures substantially improves mitochondrial energy metabolism and hepatic differentiation of human Liver Cell lines. This practice-changing culture method may prove to prolong the in-vitro maintenance of primary hepatocytes and increase hepatic differentiation of stem Cells.
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amc bio artificial Liver culturing enhances mitochondrial biogenesis in human Liver Cell lines the role of oxygen medium perfusion and 3d configuration
Mitochondrion, 2017Co-Authors: Aziza A. A. Adam, Perry D. Moerland, Ronald Oude P J Elferink, Aldo Jongejan, Robert A F M Chamuleau, Martien Van Wenum, Vincent A Van Der Mark, Riekelt H Houtkooper, Ronald J A Wanders, Ruurdtje HoekstraAbstract:Abstract Background Human Liver Cell lines, like HepaRG and C3A, acquire higher functionality when cultured in the AMC-Bio-Artificial Liver (AMC-BAL). The three main differences between BAL and monolayer culture are the oxygenation (40% vs 20%O2), dynamic vs absent medium perfusion and 3D vs 2D configuration. Here, we investigated the background of the differences between BAL-cultures and monolayers. Methods We performed whole-genome microarray analysis on HepaRG monolayer and BAL-cultures. Next, mitochondrial biogenesis was studied in monolayer and BAL-cultures of HepaRG and C3A. The driving forces for mitochondrial biogenesis by BAL-culturing were investigated in representative culture models differing in oxygenation level, medium flow or 2D vs 3D configuration. Results Gene-sets related to mitochondrial energy metabolism were most prominently up-regulated in HepaRG-BAL vs monolayer cultures. This was confirmed by a 2.4-fold higher mitochondrial abundance with increased expression of mitochondrial OxPhos complexes. Moreover, the transcript levels of mitochondria-encoded genes were up to 3.6-fold induced and mitochondrial membrane potential activity was 8.3-fold increased in BAL vs monolayers. Culturing with 40% O2, dynamic medium flow and/or in 3D increased the mitochondrial abundance and expression of mitochondrial complexes vs standard monolayer culturing. The stimulatory effect of the BAL culture on mitochondrial biogenesis was confirmed in C3A Cells in which mitochondrial abundance increased 2.2-fold with induction of mitochondria-encoded genes. Conclusions and general significance The increased functionality of Liver Cell lines upon AMC-BAL culturing is associated with increased mitochondrial biogenesis. High oxygenation, medium perfusion and 3D configuration contribute to the up-regulation of the mitochondrial biogenesis.
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evaluation of a new immortalized human fetal Liver Cell line cbal111 for application in bioartificial Liver
Journal of Hepatology, 2008Co-Authors: Paul P C Poyck, Tessa V Van Der Hoeven, Albert C W A Van Wijk, Ronald Oude P J Elferink, Dirk R De Waart, Thomas M. Gulik, Robert A F M Chamuleau, Ruurdtje HoekstraAbstract:Background/Aims Clinical use of bioartificial Livers (BAL) relies heavily on the development of human Liver Cell lines. The aim of this study was to assess the potential of the recently developed human fetal Liver Cell line cBAL111 for application in the AMC-BAL. Methods Laboratory-scale AMC-BAL bioreactors were loaded with 20 or 200 million cBAL111 Cells and were cultured for 3days. Parameters for hepatocyte-specific function and general metabolism were determined daily using tests with culture medium or 100% human serum. The bioreactors were also analyzed for mRNA levels of Liver-specific genes and histology. Results cBAL111 eliminated ammonia at a rate up to 49% of that in primary porcine hepatocytes (PPH), despite a low (1.1%) urea production. Transcript levels of glutamine synthetase (GS) were 570% of that in human Liver, whereas genes of the urea cycle showed low expression. GS expression was confirmed immunohistochemically, and glutamine was produced by the Cells. cBAL111 eliminated galactose (90.1% of PPH) and lidocaine (0.1% of PPH) and produced albumin (6% of PPH). Human serum did not increase function of cBAL111. Conclusions cBAL111 showed Liver-specific functionality when cultured inside the AMC-BAL and eliminated ammonia mainly by the activity of GS, and not through the urea cycle.