scholarly journals Delineating Biological Pathways Unique to Embryonic Stem Cell-Derived Insulin-Producing Cell Lines from Their Noninsulin-Producing Progenitor Cell Lines

Endocrinology ◽  
2010 ◽  
Vol 151 (8) ◽  
pp. 3600-3610
Author(s):  
Tian Sheng Chen ◽  
Soon Sim Tan ◽  
Ronne Wee Yeh Yeo ◽  
Bao Ju Teh ◽  
Ruihua Luo ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Signaling Pathway ◽  
Cell Lines ◽  
Progenitor Cell ◽  
Embryonic Stem ◽  
Biochemical Pathways ◽  
Cellular Assays ◽  
Ppar Signaling

To identify unique biochemical pathways in embryonic stem cell-derived insulin-producing cells as potential therapeutic targets to prevent or delay β-cell dysfunction or death in diabetic patients, comparative genome-wide gene expression studies of recently derived mouse insulin-producing cell lines and their progenitor cell lines were performed using microarray technology. Differentially expressed genes were functionally clustered to identify important biochemical pathways in these insulin-producing cell lines. Biochemical or cellular assays were then performed to assess the relevance of these pathways to the biology of these cells. A total of 185 genes were highly expressed in the insulin-producing cell lines, and computational analysis predicted the pentose phosphate pathway (PPP), clathrin-mediated endocytosis, and the peroxisome proliferator-activated receptor (PPAR) signaling pathway as important pathways in these cell lines. Insulin-producing ERoSHK cells were more resistant to hydrogen peroxide (H2O2)-induced oxidative stress. Inhibition of PPP by dehydroepiandrosterone and 6-aminonicotinamide abrogated this H2O2 resistance with a concomitant decrease in PPP activity as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Clathrin-mediated endocytosis, which is essential in maintaining membrane homeostasis in secreting cells, was up-regulated by glucose in ERoSHK but not in their progenitor ERoSH cells. Its inhibition by chlorpromazine at high glucose concentration was toxic to the cells. Troglitazone, a PPARG agonist, up-regulated expression of Ins1 and Ins2 but not Glut2. Gene expression analysis has identified the PPP, clathrin-mediated endocytosis, and the PPAR signaling pathway as the major delineating pathways in these insulin-producing cell lines, and their biological relevance was confirmed by biochemical and cellular assays.

Characterization and Gene Expression Profiling of Five New Human Embryonic Stem Cell Lines Derived in Taiwan

2006 ◽  
Vol 15 (4) ◽  
pp. 532-555 ◽  
Author(s):  
Steven Shoei-Lung Li ◽  
Yung-Hsien Liu ◽  
Chao-Neng Tseng ◽  
Tung-Liang Chung ◽  
Tzi-Yi Lee ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Gene Expression Profiling ◽  
Cell Lines ◽  
Expression Profiling ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Stem Cell Lines

scholarly journals Embryonic stem cell microenvironment enhances proliferation of human retinal pigment epithelium cells by activating the PI3K signaling pathway

2020 ◽  
Author(s):  
Jiahui Liu ◽  
Liu Yang ◽  
Xiaoran Wang ◽  
Shoubi Wang ◽  
Zheqian Huang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Retinal Pigment Epithelium ◽  
Embryonic Stem Cell ◽  
Signaling Pathway ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Embryonic Stem ◽  
Pi3k Signaling ◽  
Pi3k Signaling Pathway

Abstract Purpose This study aimed to investigate whether the mouse embryonic stem cell (ESC) microenvironment improves the stem cell phenotype and proliferation properties of human retinal pigment epithelium (hRPE) cells by regulating the PI3K signaling pathway. Methods Primary hRPE cells were cocultured with either ESCs or human corneal epithelial cells (CECs) for 72 hours, after which their proliferation, apoptosis, cell cycle progression, and colony formation were assayed to evaluate changes in their biological characteristics. Gene expression was detected by real-time PCR and protein levels were determined by western blotting or immunofluorescence. LY294002, an antagonist of the PI3K signaling pathway, was used to further confirm the mechanism involved. Results In comparison to hRPE cells cultured alone, hRPE cells cocultured with ESCs had an increased proliferative capacity, reduced apoptotic rate, and higher colony-forming efficiency. The expression of the stem cell-associated marker KLF4 and the differentiation marker CRALBP increased and decreased, respectively, in hRPE cells isolated from the ESC coculture. Furthermore, PI3K pathway-related genes were significantly up-regulated in hRPE cells after exposure to ESCs. LY294002 reversed the pro-proliferative effect of ESCs on hRPE cells. In contrast, CECs did not share the ability of ESCs to influence the biological behavior and gene expression of hRPE cells. Conclusions Our findings indicate that the ESC microenvironment enhances stemness and proliferation of hRPE cells, partially via activation of the PI3K signaling pathway. This study may have a significant impact and clinical implication on cell therapy in regenerative medicine, specifically for age-related macular degeneration.

scholarly journals Embryonic stem cell microenvironment enhances proliferation of human retinal pigment epithelium cells by activating the PI3K signaling pathway

2020 ◽  
Author(s):  
Jiahui Liu ◽  
Liu Yang ◽  
Xiaoran Wang ◽  
Shoubi Wang ◽  
Zheqian Huang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Retinal Pigment Epithelium ◽  
Embryonic Stem Cell ◽  
Signaling Pathway ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Embryonic Stem ◽  
Pi3k Signaling ◽  
Pi3k Signaling Pathway

Abstract Purpose. This study aimed to investigate whether the mouse embryonic stem cell (ESC) microenvironment improves the stem cell phenotype and proliferation properties of human retinal pigment epithelium (hRPE) cells by regulating the PI3K signaling pathway. Methods. Primary hRPE cells were cocultured with either ESCs or human corneal epithelial cells (CECs) for 72 hours, after which their proliferation, apoptosis, cell cycle progression, and colony formation were assayed to evaluate changes in their biological characteristics. Gene expression was detected by real-time PCR and protein levels were determined by western blotting or immunofluorescence. LY294002, an antagonist of the PI3K signaling pathway, was used to further confirm the mechanism involved. Results. In comparison to hRPE cells cultured alone, hRPE cells cocultured with ESCs had an increased proliferative capacity, reduced apoptotic rate, and higher colony-forming efficiency. The expression of the stem cell-associated marker KLF4 and the differentiation marker CRALBP increased and decreased, respectively, in hRPE cells isolated from the ESC coculture. Furthermore, PI3K pathway-related genes were significantly up-regulated in hRPE cells after exposure to ESCs. LY294002 reversed the pro-proliferative effect of ESCs on hRPE cells. In contrast, CECs did not share the ability of ESCs to influence the biological behavior and gene expression of hRPE cells. Conclusions. Our findings indicate that the ESC microenvironment enhances stemness and proliferation of hRPE cells, partially via activation of the PI3K signaling pathway. This study may have a significant impact and clinical implication on cell therapy in regenerative medicine, specifically for age-related macular degeneration.

scholarly journals Establishment of Mouse Embryonic Stem Cell-Derived Erythroid Progenitor Cell Lines Able to Produce Functional Red Blood Cells

PLoS ONE ◽  
2008 ◽  
Vol 3 (2) ◽  
pp. e1544 ◽  
Author(s):  
Takashi Hiroyama ◽  
Kenichi Miharada ◽  
Kazuhiro Sudo ◽  
Inaho Danjo ◽  
Naoko Aoki ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Red Blood Cells ◽  
Cell Lines ◽  
Blood Cells ◽  
Progenitor Cell ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Erythroid Progenitor ◽  
Erythroid Progenitor Cell
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