Journal: Frontiers in Veterinary Science

Article Title: Next-Generation Intestinal Toxicity Model of Human Embryonic Stem Cell-Derived Enterocyte-Like Cells

doi: 10.3389/fvets.2021.587659

Figure Lengend Snippet: Differentiation of human embryonic stem cells into human enterocytes. (A) Scheme of the differentiation protocol of human embryonic stem cell-derived enterocyte-like cells (hESC-ELCs) from hESCs. (B) Representative morphological changes during differentiation from hESCs to definitive endoderm (DE), hindgut (HG), and hESC-ELCs. Scale bars , 100 μm. (C) Expression levels of the intestine-specific marker ( CDX2 ), enterocyte markers ( VIL1 and SI ), and the tight junction markers ( ZO-1, OCLN, CLDN1, CLDN3 , and CLDN5 ) in hESCs and hESC-ELCs assessed by qPCR analysis. Data are presented as the mean ± SEM. * p < 0.05; ** p < 0.01. (D) Immunofluorescent staining of hESCs and hESC-ELCs with the intestine-specific marker ( CDX2 ) and enterocyte marker ( VIL1 ). Scale bars , 20 μm. (E) Gene Ontology (GO) network and Enrichment Pathway analysis for the hESC-ELCs assessed by ClueGO. Upregulated differentially expressed genes (DEGs; fold change > 128) in hESC-ELCs compared to H9 hESCs were analyzed. (F) The GOs of DEGs in (E) were presented with their p values.

Article Snippet: From the National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO), the RNA-seq data of H9 hESCs (GEO no. GSE118106), differentiated Caco-2 cells (GEO no. GSE97977), Hutu-80 cells (GEO no. GSE59857), human fetal small intestine (GEO no. GSE108369), and human adult small intestine (GEO no. GSE117875) were obtained.

Techniques: Derivative Assay, Expressing, Marker, Staining

Journal: Frontiers in Veterinary Science

Article Title: Next-Generation Intestinal Toxicity Model of Human Embryonic Stem Cell-Derived Enterocyte-Like Cells

doi: 10.3389/fvets.2021.587659

Figure Lengend Snippet: Transcriptomic assessment of human embryonic stem cell-derived enterocyte-like cells (hESC-ELCs) and other small intestine models. hESC-ELCs, rat small intestine, Caco-2 cells, Hutu-80 cells, and human adult and fetal small intestines without any treatment were used for RNA sequencing (RNA-seq). (A) Heatmap of a total number of 10,020 genes used to cluster the models hierarchically. The heatmap represents six models ( columns ) and their genes ( rows ). The color scale at the top represents the expression level, where red, blue , and white indicate upregulation, downregulation, and an unaltered expression, respectively. (B) Relative distances of the seven models visualized using principal component analysis (PCA). Dots with colors represent each model, and the red arrows are for each gene in the transcriptomic analysis. Clustered samples share the colors and lines . hEscElc , hESC-ELCs; rSI , rat small intestine; Caco2 , Caco-2 cells; Hutu80 , Hutu-80 cells; hFetalSI , human fetal small intestine; hAdultSI , human adult small intestine. (C) Venn diagram visualizing the intersection between hESC-ELCs and the in vivo models. The intersection of the human and rat adult small intestines has 214 genes in common. (D) The enriched biological pathways were assessed by ClueGO. The intersection of the human adult and fetal small 698 intestines has 145 genes in common. (E) The enriched pathways were also assessed. The intersection of hESC-ELCs and the human adult small intestines has 46 genes in common. (F) The enriched top 28 pathways were assessed by DAVID. (G) Comparison of the hESC-ELCs and human adult small intestines in intestine-related biological pathways.

Article Snippet: From the National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO), the RNA-seq data of H9 hESCs (GEO no. GSE118106), differentiated Caco-2 cells (GEO no. GSE97977), Hutu-80 cells (GEO no. GSE59857), human fetal small intestine (GEO no. GSE108369), and human adult small intestine (GEO no. GSE117875) were obtained.

Techniques: Derivative Assay, RNA Sequencing, Expressing, In Vivo, Comparison

Journal: Frontiers in Veterinary Science

Article Title: Next-Generation Intestinal Toxicity Model of Human Embryonic Stem Cell-Derived Enterocyte-Like Cells

doi: 10.3389/fvets.2021.587659

Figure Lengend Snippet: Relative mRNA expression levels in rat small intestines, Caco-2 cells, Hutu-80 cells, and human embryonic stem cell-derived enterocyte-like cells (hESC-ELCs). Rat small intestines, Caco-2 cells, Hutu-80 cells, and hESC-ELCs were treated with chloramphenicol (CHL), cycloheximide (CHX), cytarabine (Ara-C), diclofenac (DIC), fluorouracil (5-FU), indomethacin (INDO), methotrexate (MTX), oxytetracycline (OTC), or vehicle and assessed by quantitative PCR (qPCR). (A) The relative expressions of hCYP2C9/rCyp2c11, hCYP2C19/rCyp2c6v1, hCYP2D6/yCyp2d3, hCYP2E1/rCyp2e1, hCYP3A4/rCyp3a2, MAOA, NAT2, HO1, IL1RN, TNF , and hCYP24A1/rCyp24a1 were monitored. Also, the ratio of Bax / Bcl-2 was calculated. Data shown are the mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001. (B) Heatmap of the entire set of genes represented by four models ( columns ) and their genes treated with each drug ( rows ). The color scale at the top represents the expression level, where red, blue , and white indicate upregulation, downregulation, and an unaltered expression, respectively. (C) Relative distances of the four models visualized using principal component analysis (PCA). Black dots represent each model, and the red arrows are for each gene in the qPCR analysis. hEscElc , hESC-ELCs; rSI , rat small intestine; Caco2 , Caco-2 cells; Hutu80 , Hutu-80 cells. (D–F) Correlations with the in vivo model under treatment of intestinal toxicants of hESC-ELCs (D) , Caco-2 cells (E) , and Hutu-80 cells (F) (for all panels, the x -axis shows the rat in vivo gene expression and the y -axis shows the cell gene expression). The strength of the correlations between the in vitro models and the in vivo model is shown as Pearson's correlation coefficient ( r ) with associated p -values.

Article Snippet: From the National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO), the RNA-seq data of H9 hESCs (GEO no. GSE118106), differentiated Caco-2 cells (GEO no. GSE97977), Hutu-80 cells (GEO no. GSE59857), human fetal small intestine (GEO no. GSE108369), and human adult small intestine (GEO no. GSE117875) were obtained.

Techniques: Expressing, Derivative Assay, Real-time Polymerase Chain Reaction, In Vivo, Gene Expression, In Vitro

Journal: Frontiers in Veterinary Science

Article Title: Next-Generation Intestinal Toxicity Model of Human Embryonic Stem Cell-Derived Enterocyte-Like Cells

doi: 10.3389/fvets.2021.587659

Figure Lengend Snippet: Representative morphological changes in human embryonic stem cell-derived enterocyte-like cells (hESC-ELCs) and in the other models under treatment of intestinal toxicants. Rat small intestines, Caco-2 cells, Hutu-80 cells, and hESC-ELCs were treated with chloramphenicol (CHL), cycloheximide (CHX), cytarabine (Ara-C), diclofenac (DIC), fluorouracil (5-FU), indomethacin (INDO), methotrexate (MTX), oxytetracycline (OTC), or vehicle and the morphological changes were analyzed. (A) The rat tissues were stained with hematoxylin and eosin (HandE), and the rat tissues and cells were assessed. Arrows indicate the damaged intestinal architectures in the rat intestines, and arrowheads indicate the characteristics of apoptotic cell death in Caco-2 cells, Hutu-80 cells, and hESC-ELCs. All scale bars , 100 μm. (B–E) Apoptotic indices were also calculated in rat small intestines (B) , Caco-2 cells (C) , Hutu-80 cells (D) , and hESC-ELCs (E) . Data shown are the mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: From the National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO), the RNA-seq data of H9 hESCs (GEO no. GSE118106), differentiated Caco-2 cells (GEO no. GSE97977), Hutu-80 cells (GEO no. GSE59857), human fetal small intestine (GEO no. GSE108369), and human adult small intestine (GEO no. GSE117875) were obtained.

Techniques: Derivative Assay, Staining

Journal: Stem Cell Research & Therapy

Article Title: Advances in retinal pigment epithelial cell transplantation for retinal degenerative diseases

doi: 10.1186/s13287-024-04007-5

Figure Lengend Snippet: Summary of recent completed and ongoing clinical trials involving transplantation of RPE in patients with diagnosed eye ailments

Article Snippet: Dry AMD: 16 patients (age 69–85 years) with cohort 1; BCVA ≤ 20/200, cohort 2: 20/80 to 20/400 , NCT02590692 (Unknow status, phase 1/2a, 3-year median follow-up) , Regenerative Patch Technologies, LLC , Cell source: hESC (H9) derived RPE, allogenic Form: cell monolayer with parylene C substrate , Tacrolimus started 8 days before surgery, continued to day 42, and gradually reduced until day 60 , One patient could not be transplanted Patients in cohort 1 had subretinal haemorrhage, retinal or macular edema, focal retinal detachment, or RPE detachment, which was mitigated in cohort 2 with an improved haemostasis during surgery , Implanted eyes improved by > 5 letters BCVA throughout median 3-year follow-up Safe and well-tolerant in all patients 3 patients had fixation detected over the transplant in cohort 1 , [ , , 83] .

Techniques: Transplantation Assay, Derivative Assay, Suspension, Membrane, Migration, Stripping Membranes, Mutagenesis

Journal: Stem Cells International

Article Title: Human Very Small Embryonic-Like Stem Cells Are Present in Normal Peripheral Blood of Young, Middle-Aged, and Aged Subjects

doi: 10.1155/2016/7651645

Figure Lengend Snippet: Relative expressions of PSC markers, Oct-4, Nanog, and Sox2, were measured by RT-qPCR and compared using equal amounts of mRNA isolated from CNT PB of young, middle, and older healthy volunteers. The relative expression of each PSC marker is calculated according to a positive control PCR (RNA isolated from H9 and HUES 3 hESC lines). Data represent the mean ± standard deviation for each age group. The difference in mRNA expression of these markers between the three groups of age was not statistically significant ( P > 0.05, Kruskal-Wallis test).

Article Snippet: RNA isolated from H9 and HUES 3 hESC lines [ , ], kindly provided by the “Plateforme Cellules Souches Embryonnaires Humaines” (Inserm U602 Villejuif, France), was used as reference sample for each PCR reaction.

Techniques: Quantitative RT-PCR, Isolation, Expressing, Marker, Positive Control, Standard Deviation