Journal: Acta biomaterialia

Article Title: Probing prodrug metabolism and reciprocal toxicity with an integrated and humanized multi-tissue organ-on-a-chip platform

doi: 10.1016/j.actbio.2020.02.015

Figure Lengend Snippet: A summary of each organoid or tissue construct type with the cells making them up. Also described are the various cell sources and cell types (primary, cell line, iPSC-derived, etc.), relative percentage in each model, and documented tissue specific functionalities.

Article Snippet: Organoid or tissue construct type Cell types cell source Type of cell Percentage in model Documented functionality Citation Liver Hepatocytes Bioreclamation Primary human 75% Albumin and urea secretion; Drug metabolism; Toxicity responses [ 18 , 47 ] Stellate cells Bioreclamation Primary human 10% Kupffer cells Sekisui XenoTech Primary human 10% Liver-Derived endothelial cells Lonza primary human 5% Cardiac Cardiomyocytes Ncardia iPSC 75% Spontaneous beating; Beating changes in response to drugs; Toxicity responses [ 18 , 47 ] Cardiac fibroblasts ScienCell Primary HUMAN 20% Cardiac endothelial cells ScienCell Primary human 5% Lung (3-tissue system) A549 Alveolar epithelial cells Lonza Human cell line 100% n/a – Lung (6-tissue system) Lung fibroblasts Lonza Primary human 20% n/a – Human bronchial epithelial cells Lonza Primary human 80% Endothelial HUVEC Lonza Primary human 100% n/a – Testis Spermatogonial stem cells Donor via NDRI Primary human ~80% Androgen production; Supporting germ cell differentiation; Toxicity responses; Virus replication [ 34 – 36 ] Leydig cells Donor via NDRI Primary human ~10% Sertoli cells Donor via NDRI Primary human ~10% Peritubular cells Donor via NDRI Primary human Contaminant of SSC populations Brain Brain microvascular endothelial cells Cell Systems Primary human 30% Barrier function to the middle of the spheroid [ 37 ] Brain vascular pericytes ScienCell Primary human 15% Astrocytes ScienCell Primary human 15% Microglia Tempo Bioscience IPSC 5% Oligodendrocytes Tempo Bioscience Oligodendrocyte progenitor cells 15% Neural cells Axol Biosciences Neural stem cells 20% Open in a separate window A summary of each organoid or tissue construct type with the cells making them up.

Techniques: Construct, Cell Differentiation, Virus

Journal: Acta biomaterialia

Article Title: Probing prodrug metabolism and reciprocal toxicity with an integrated and humanized multi-tissue organ-on-a-chip platform

doi: 10.1016/j.actbio.2020.02.015

Figure Lengend Snippet: Drug toxicity assessment of ifosfamide in a 6-organoid system. L/D imaging of liver, cardiac, lung, endothelial, brain, and testis organoids under Control (no drug, a–c), Condition 1 (with ifosfamide and liver organoid, d–f), and Condition 2 (with ifosfamide and without liver organoid, g and h). In Condition 1, the metabolized drug caused downstream toxicity in the brain organoid; in Condition 2, no significant toxicity occurred. Green stain: calcein AM-stained viable cells; Red stain: ethidium homodimer 1-stained dead cells. Scale bar represents 100 μM. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Organoid or tissue construct type Cell types cell source Type of cell Percentage in model Documented functionality Citation Liver Hepatocytes Bioreclamation Primary human 75% Albumin and urea secretion; Drug metabolism; Toxicity responses [ 18 , 47 ] Stellate cells Bioreclamation Primary human 10% Kupffer cells Sekisui XenoTech Primary human 10% Liver-Derived endothelial cells Lonza primary human 5% Cardiac Cardiomyocytes Ncardia iPSC 75% Spontaneous beating; Beating changes in response to drugs; Toxicity responses [ 18 , 47 ] Cardiac fibroblasts ScienCell Primary HUMAN 20% Cardiac endothelial cells ScienCell Primary human 5% Lung (3-tissue system) A549 Alveolar epithelial cells Lonza Human cell line 100% n/a – Lung (6-tissue system) Lung fibroblasts Lonza Primary human 20% n/a – Human bronchial epithelial cells Lonza Primary human 80% Endothelial HUVEC Lonza Primary human 100% n/a – Testis Spermatogonial stem cells Donor via NDRI Primary human ~80% Androgen production; Supporting germ cell differentiation; Toxicity responses; Virus replication [ 34 – 36 ] Leydig cells Donor via NDRI Primary human ~10% Sertoli cells Donor via NDRI Primary human ~10% Peritubular cells Donor via NDRI Primary human Contaminant of SSC populations Brain Brain microvascular endothelial cells Cell Systems Primary human 30% Barrier function to the middle of the spheroid [ 37 ] Brain vascular pericytes ScienCell Primary human 15% Astrocytes ScienCell Primary human 15% Microglia Tempo Bioscience IPSC 5% Oligodendrocytes Tempo Bioscience Oligodendrocyte progenitor cells 15% Neural cells Axol Biosciences Neural stem cells 20% Open in a separate window A summary of each organoid or tissue construct type with the cells making them up.

Techniques: Imaging, Staining

Journal: Acta biomaterialia

Article Title: Probing prodrug metabolism and reciprocal toxicity with an integrated and humanized multi-tissue organ-on-a-chip platform

doi: 10.1016/j.actbio.2020.02.015

Figure Lengend Snippet: Viability quantification under insult by ifosfamide in a 6-organoid system. Quantification of live cell ratios (L/L + D) for liver, cardiac, lung, endothelial, brain, and testis organoids under Control (no drug), Condition 1 (drug with liver organoid), and Condition 2 (drug without liver organoid). A significant reduction in viability is observed for the brain organoid under Condition 1 (red). Statistical significance: ** p < 0.05. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Organoid or tissue construct type Cell types cell source Type of cell Percentage in model Documented functionality Citation Liver Hepatocytes Bioreclamation Primary human 75% Albumin and urea secretion; Drug metabolism; Toxicity responses [ 18 , 47 ] Stellate cells Bioreclamation Primary human 10% Kupffer cells Sekisui XenoTech Primary human 10% Liver-Derived endothelial cells Lonza primary human 5% Cardiac Cardiomyocytes Ncardia iPSC 75% Spontaneous beating; Beating changes in response to drugs; Toxicity responses [ 18 , 47 ] Cardiac fibroblasts ScienCell Primary HUMAN 20% Cardiac endothelial cells ScienCell Primary human 5% Lung (3-tissue system) A549 Alveolar epithelial cells Lonza Human cell line 100% n/a – Lung (6-tissue system) Lung fibroblasts Lonza Primary human 20% n/a – Human bronchial epithelial cells Lonza Primary human 80% Endothelial HUVEC Lonza Primary human 100% n/a – Testis Spermatogonial stem cells Donor via NDRI Primary human ~80% Androgen production; Supporting germ cell differentiation; Toxicity responses; Virus replication [ 34 – 36 ] Leydig cells Donor via NDRI Primary human ~10% Sertoli cells Donor via NDRI Primary human ~10% Peritubular cells Donor via NDRI Primary human Contaminant of SSC populations Brain Brain microvascular endothelial cells Cell Systems Primary human 30% Barrier function to the middle of the spheroid [ 37 ] Brain vascular pericytes ScienCell Primary human 15% Astrocytes ScienCell Primary human 15% Microglia Tempo Bioscience IPSC 5% Oligodendrocytes Tempo Bioscience Oligodendrocyte progenitor cells 15% Neural cells Axol Biosciences Neural stem cells 20% Open in a separate window A summary of each organoid or tissue construct type with the cells making them up.

Techniques:

Journal: Journal of the American College of Cardiology

Article Title: Cardioprotective Effects of HSP72 Administration on Ischemia-Reperfusion Injury

doi: 10.1016/j.jacc.2017.07.762

Figure Lengend Snippet: (A) Human primary cardiomyocytes were exposed to 2.6 mM hydrogen peroxide (H2O2) at time T = 0 h and throughout the course of the study. Treatment of cells 30 min after the start of intoxication with 3E10-Fv alone ( ) or HSP72 alone ( ) did not affect the increase in cell death, while treatment with Fv-HSP72 ( ) significantly attenuated apoptosis (p = 0.0006). Addition of 3E10-Fv prior to Fv-HSP72 treatment ( ) inhibited Fv-HSP72 efficacy, but still significantly attenuated apoptosis at 12 h (p = 0.002). (B) To study cardiomyocyte exposure overnight, cells were intoxicated with 1.4 mM H2O2 at time T = 0 h and throughout the course of the study. Fv-HSP72 treatment at 30 min after the start of intoxication significantly reduced the percentage of cell death at 17 h compared to the H2O2-only control (p = 0.0003), while treatment with 3E10-Fv alone or HSP72 alone showed no statistically significant inhibition. Percentages reflect normalization of the average fluorescent signal at the last reading divided by the maximum signal obtained after total cell lysis in 3 of the No H2O2 control wells. Number of wells receiving each treatment cited in parentheses. *statistically significant; ***highly significant. Error bars represent SE of the mean. Abbreviations as in Figure 1.

Article Snippet: Human primary cardiomyocytes (Cat. #6200, ScienCell, Carlsbad, California) were thawed and the 1 ml (1 × 10 6 cells/vial) suspended in 19 ml of phenol-red free cardiac myocyte media (includes 5% fetal bovine serum, 1% cardiac myocyte growth supplement, 1X penicillin/streptomycin; Cat. #6201-prf, ScienCell) prior to seeding into white, opaque bottom 96-well tissue culture plates at 5,000 cells per well (Cat. #3917, Corning Life Sciences, Tewksbury, Massachusetts).

Techniques: Control, Inhibition, Lysis

Journal: Journal of the American College of Cardiology

Article Title: Cardioprotective Effects of HSP72 Administration on Ischemia-Reperfusion Injury

doi: 10.1016/j.jacc.2017.07.762

Figure Lengend Snippet: A single-chain variable fragment (Fv) of the 3E10 antibody transports heat shock protein (HSP)72 into cardiomyocytes through the equilibrative nucleoside transporter 2 (ENT2) channel, where the HSP inhibited apoptosis in a rabbit model of ischemia-reperfusion injury. Either moiety alone is not cardioprotective.

Article Snippet: Human primary cardiomyocytes (Cat. #6200, ScienCell, Carlsbad, California) were thawed and the 1 ml (1 × 10 6 cells/vial) suspended in 19 ml of phenol-red free cardiac myocyte media (includes 5% fetal bovine serum, 1% cardiac myocyte growth supplement, 1X penicillin/streptomycin; Cat. #6201-prf, ScienCell) prior to seeding into white, opaque bottom 96-well tissue culture plates at 5,000 cells per well (Cat. #3917, Corning Life Sciences, Tewksbury, Massachusetts).

Techniques:

Journal: European journal of immunology

Article Title: Sca-1 + cardiac fibroblasts promote development of heart failure

doi: 10.1002/eji.201847583

Figure Lengend Snippet: Sca-1+ cardiac fibroblasts are potent GM-CSF-producers during cardiac inflammation. Cardiac fibroblasts were harvested from wild type BALB/c mice with EAM on days 0, 14 or 21 for flow cytometry analysis. (A) Representative flow cytometry plots depict gating strategies for cardiac fibroblast subsets. (B) Flow cytometric quantification of the numbers of cardiac fibroblast subsets per mg cardiac tissue at different time points after EAM induction. (n = 4 for EAM day 0 group; n = 6 for EAM day 14 group; n = 7 for EAM day 21 group; CF cardiac fibroblast) (C) Demonstration of cell number changes in cardiac fibroblast subsets after EAM induction. (D) Representative flow cytometry= plots showing GM-CSF staining of cardiac fibroblast subsets on day 21 of EAM. (E and F) Representative t-SNE clustering plots of (E) cardiac fibroblast subsets and GM-CSF-positive cardiac fibroblasts or (F) all GM-CSF-positive cardiac cells on day 21 of EAM using t-SNE dimensionality reduction algorithm plugin in FlowJo 10.4.2 (FlowJo, LLC). (G) Relative frequency of different cell populations out of all GM-CSF-positive cells in the hearts on day 21 of EAM. (H) Frequency and number of GM-CSF-positive Sca-1+ cardiac fibroblasts per mg cardiac tissue in immunized mice (n = 9) versus mock-immunized mice (n = 10) on day 21 of EAM. (I) Frequency and number of GM-CSF-positive Sca-1+ cardiac fibroblasts per mg cardiac tissue in immunized WT mice and Il17ra−/− mice on day 21 of EAM (n = 10 per group). Data are representative of three independent experiments (A–I). Groups were compared using one-way ANOVA followed by Tukey’s post-test (B) or Student’s t-test (H–I). *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: Human primary cardiac fibroblasts We acquired human primary cardiac fibroblasts from Cell Applications, Inc (San Diego, CA).

Techniques: Flow Cytometry, Staining

Journal: European journal of immunology

Article Title: Sca-1 + cardiac fibroblasts promote development of heart failure

doi: 10.1002/eji.201847583

Figure Lengend Snippet: Sca-1+ cardiac fibroblasts are potent producers of different cytokines and chemokines and are plastic in response to environment changes. (A) The mRNA expression levels of Csf2, Ccl2, Cxcl10, Ccl11 and Postn in na¨ıve mouse heart homogenates, EAM day 21 mouse heart homogenates and FACS-sorted Sca-1+ and Sca-1− cardiac fibroblasts from EAM day 21 mouse hearts (n =5 per group; n.d. not detected; WH = whole heart homogenate). Expression levels were analyzed by qPCR (2− ΔΔ Ct values relative to Gapdh expression levels and=na¨ıve whole heart transcripts). (B) The mRNA expression levels of Csf2, Ccl2, Cxcl10 and Ccl11 in in vitro cultured Sca-1+ cardiac fibroblasts stimulated with IL-17A, IL-13, IFN-γ, or unstimulated (control). Expression levels were analyzed by qPCR (2−ΔΔCt values relative to Gapdh expression levels and unstimulated controls). (C) Representative histogram of mCherry (CCL2) expression in cultured Sca-1+ cardiac fibroblasts under Th17 conditions or unstimulated. (D) Schematic of the experiment procedures for all data showing in E–F. (E) Expression levels of Ccl2, Csf2 and Ccl11 mRNA in cells harvested on day 3 or day 6. Expression levels were analyzed by qPCR (2−͉ ΔΔCt values relative to Gapdh expression levels and unstimulated controls). (F) Protein concentrations of secreted CCL11 and GM-CSF in the culture supernatant harvested on day 3 or day 6 were measured by quantitative sandwich ELISA. Data are shown as mean + SD of five mice per group (A) or of technical triplicates (B, E–F). Data are representative of three indepen dent experiments (A–F). Groups were compared using one-way ANOVA followed by Tukey’s post-test (A, E–F) or Student’s t-test (B). **p < 0.01; ***p < 0.001.

Article Snippet: Human primary cardiac fibroblasts We acquired human primary cardiac fibroblasts from Cell Applications, Inc (San Diego, CA).

Techniques: Expressing, In Vitro, Cell Culture, Control, Sandwich ELISA

Journal: European journal of immunology

Article Title: Sca-1 + cardiac fibroblasts promote development of heart failure

doi: 10.1002/eji.201847583

Figure Lengend Snippet: IL-17A induces GM-CSF production in Sca-1+ cardiac fibroblasts through synergistic activation of NF-κB and NFAT2. (A–C) Sca-1+ cardiac fibroblasts were MACS-sorted from naïve WT mouse hearts and cultured under Th17 conditions (IL-17A TNF-α), Th2 conditions (IL-13) or unstimulated (control) for two days (A) or for multiple time points (B–C). (A) Representative flow cytometry plots showing phosphorylated p-65 (pp65)-positive and/or NFAT2-positive Sca-1+ cardiac fibroblasts and representative histogram showing their GM-CSF expression. (B) Immunoblots showing the detection of total or phosphorylated Stat6, Stat3, p65 and Erk1/2 in whole cell lysate. (C) Immunoblots showing the detection of p65, Stat6, Parp1 (nuclear positive control) and pro-Caspase 3 (nuclear negative control) in the nuclear fraction of Sca-1 cardiac fibroblasts with/without stimulation. Data are representative of three independent experiments (A–C).

Article Snippet: Human primary cardiac fibroblasts We acquired human primary cardiac fibroblasts from Cell Applications, Inc (San Diego, CA).

Techniques: Activation Assay, Cell Culture, Control, Flow Cytometry, Expressing, Western Blot, Positive Control, Negative Control

Journal: European journal of immunology

Article Title: Sca-1 + cardiac fibroblasts promote development of heart failure

doi: 10.1002/eji.201847583

Figure Lengend Snippet: IL-17A signaling ablation leads to less GM-CSF-positive Sca-1+ cardiac fibroblasts and protects mice from post-MI heart failure and death. (A) Representative flow cytometry plots showing GM-CSF- and CCL2-positive Sca-1+ cardiac fibroblasts in infarcted and sham-operated mouse hearts on day 2 post-MI. Cells were pre-gated on viable CD45−CD31−Sca-1+ cardiac cells. (B) Flow cytometric quantification of GM-CSF+, CCL2+ and GM-CSF+CCL2+ Sca-1+ cardiac fibroblasts per mg cardiac tissue of infarcted or sham-operated WT mice (n =3 per group). (C) Kaplan-Meier survival curve of WT mice (n = 8) and Il17ra−/− mice (n = 17) on day 28 post-MI. (D) images Representative of Masson’s trichrome stained histology slides of the cross-sections at the middle of the hearts from WT and Il17ra−/− mice showing the expansion of the infarct area and the dilation of the left ventricles on day 28 post-MI. (Scalebar, 1mm) (E) Calculated total infarct sizes of WT mice (n = 6) and Il17ra−/− mice (n 14) on day 28 post-MI. (F) Flow cytometric quantification of GM-CSF-positive Sca-1+ cardiac fibroblasts per mg cardiac tissue in WT and Il17ra−/−=mice on day 2 post-MI (n = 5 per group). Data are representative of three independent experiments (A–F). Groups were compared using Student’s t-test (B, E–F) or Long-rank (Mantel-Cox) test (C). *p < 0.05; **p < 0.01.

Article Snippet: Human primary cardiac fibroblasts We acquired human primary cardiac fibroblasts from Cell Applications, Inc (San Diego, CA).

Techniques: Flow Cytometry, Staining

Journal: European journal of immunology

Article Title: Sca-1 + cardiac fibroblasts promote development of heart failure

doi: 10.1002/eji.201847583

Figure Lengend Snippet: GM-CSF-positive Sca-1+ cardiac fibroblasts express periostin post-MI and specific IL-17RA ablation in periostin+ cardiac fibroblasts protects mice from post-MI heart failure and death. (A) Representative flow cytometry plots showing Sca-1+ and GM-CSF+Sca-1+ cardiac fibroblasts and representative histograms of their GFP (periostin) expression in infarcted WT mouse hearts (gray shadowed), sham (black dashed line) and infarcted (black solid line) PostnCre/eGFP mouse hearts on day 2 post-MI. Cells were pre-gated on viable CD45−CD31−CD29+ cells. (B) Representative images of Masson’s trichrome stained histology slides of the cross-sections at the middle of the hearts from sham and infarcted PostnCre, infarcted Il17rafl/fl and infarcted PostnCreIl17rafl/fl mice on day 7 post-MI (PostnCreIl17rafl/fl mice were labeled in this Figure as PostnIL−17RAKO) showing the expansion of the infarct area and the dilation of the left ventricle of the infarcted mouse hearts. (Scalebar, 1mm) (C) Calculated total infarct sizes of infarcted Il17rafl/fl mice (n = 8) and PostnCreIl17rafl/fl mice (n =9) on day 7 post-MI. (D) Kaplan-Meier survival analysis of infarcted Il17rafl/fl mice (n = 31) and PostnCreIl17rafl/fl mice (n = 11) on day 28 post-MI. (E) Flow cytometric quantification of GM-CSF-positive Sca-1+ cardiac fibroblasts per mg cardiac tissue in PostnCre and PostnCreIl17rafl/fl mice on day 4 post-MI (n = 3 per group). (F-G) Flow cytometric quantification of the frequency (F) and the number (G) of Ly6Chi monocytes per mg cardiac tissue in PostnCre and PostnCreIl17rafl/fl mice on day 4 post-MI (n = 3 per group). “PostnIL−17RAKO” stands for “PostnCreIl17rafl/fl mice”. “CFs” stands for “cardiac fibroblasts”. Data are representative of three independent = experiments (A–G). Groups were compared using Student’s t-test (C, E–G) or Long-rank (Mantel-Cox) test (D). *p < 0.05, **p < 0.01.

Article Snippet: Human primary cardiac fibroblasts We acquired human primary cardiac fibroblasts from Cell Applications, Inc (San Diego, CA).

Techniques: Flow Cytometry, Expressing, Staining, Labeling

Journal: European journal of immunology

Article Title: Sca-1 + cardiac fibroblasts promote development of heart failure

doi: 10.1002/eji.201847583

Figure Lengend Snippet: GM-CSF-expressing cardiac fibroblast subset was found in human cardiac fibroblasts and exhibits similar functional plasticity. (A) Representative flow cytometry plots showing the gating strategies for cultured human primary cardiac fibroblasts. (B) Representative t-SNE clustering plots of three human cardiac fibroblast subsets using t-SNE dimensionality reduction algorithm plugin in FlowJo 10.4.2 (FlowJo, LLC). (C) Representative expression heat maps of cell surface markers, CD29, CD73, CD105, CD90, PDGFRα, cKit, DDR2 and GM-CSF expression in human cardiac fibroblasts using t-SNE dimensionality reduction algorithm plugin in FlowJo 10.4.2 (FlowJo, LLC). (D–E) The mRNA expression levels of CCL2, CSF2, CCL11, CCL24 and CCL26 of human primary cardiac fibroblasts cultured under IL-17A, IL-13 or unstimulated (control) for three days (D) or washed and recultured again with/without change of stimuli for another three days (E). The mRNA expression levels were analyzed by qPCR (2−ΔΔ Ct values relative to HPRT1 expression levels and unstimulated controls). Data are shown as mean SD of technical triplicates and are representative of three independent experiments (D–E). Groups were compared using Student’s t test (D) or +using one-way ANOVA followed by Tukey’s post-test (E). **p < 0.01; ***p < 0.001.

Article Snippet: Human primary cardiac fibroblasts We acquired human primary cardiac fibroblasts from Cell Applications, Inc (San Diego, CA).

Techniques: Expressing, Functional Assay, Flow Cytometry, Cell Culture, Control

Journal: European journal of immunology

Article Title: Sca-1 + cardiac fibroblasts promote development of heart failure

doi: 10.1002/eji.201847583

Figure Lengend Snippet: Human cardiac fibroblasts from heart failure patients produce GM-CSF and CCL2. (A) Representative flow cytometry plots of the concatenated sample showing gating strategy for human CD45−CD31−CD29+PDGFRα+ cardiac fibroblasts and flow cytometry plots showing GMCSF and CCL2 expressions for PDGFRα+ and PDGFRα− cardiac fibroblasts from the heart biopsies of myocarditis or ischemic heart failure patients. (B) Representative flow cytometry plots showing GM-CSF and CCL2 expression by PDGFRα+ cardiac fibroblasts from heart biopsies of five myocarditis heart failure patients. (C) Representative flow cytometry plots showing GM-CSF and CCL2 expression by PDGFRα+ cardiac fibroblasts from heart biopsies of five ischemic heart failure patients. Data are from 5 myocarditis heart failure patients and 5 ischemic heart failure patients.

Article Snippet: Human primary cardiac fibroblasts We acquired human primary cardiac fibroblasts from Cell Applications, Inc (San Diego, CA).

Techniques: Flow Cytometry, Expressing

Journal: Antioxidants (Basel, Switzerland)

Article Title: Intracellular Polyphenol Wine Metabolites Oppose Oxidative Stress and Upregulate Nrf2/ARE Pathway.

doi: 10.3390/antiox11102055

Figure Lengend Snippet: Figure 4. Dose–response effect of wine extract (WE) and sulforaphane (SFN) on oxidative stress- induced intracellular reactive oxygen species (ROS) formation. (a,b) THP-1 cells and cardiomyocytes were pre-incubated with increasing concentrations (from 200 to 800 µg/mL of total polyphenols) of WE and SFN (1–10 µM) before the addition of tert-butyl hydroperoxide (TBHP, 100 µM). Data represent the mean +/−standard deviation of measurements performed in triplicate in three different experiments; * p < 0.01 increase vs. control; ** p < 0.001 decrease vs. THBP; § p < 0.1 decrease vs. TBHP. (c) Representative flow cytometry analysis of ROS generation induced by TBHP and the inhibitory effect of the highest dose of wine extract. (d) Representative flow cytometry analysis on cell viability of the highest dose of wine extract.

Article Snippet: Human cardiomyocyte ventricular primary cells (Celprogen, Torrance, CA, USA) were cultured following the recommended manufacturer’s protocols in extracellular matrix precoated flasks and/or well plates with a ready-to-use complete growth medium (Celprogen, Torrance, CA, USA), as previously described [37].

Techniques: Incubation, Standard Deviation, Control, Cytometry

Journal: Antioxidants (Basel, Switzerland)

Article Title: Intracellular Polyphenol Wine Metabolites Oppose Oxidative Stress and Upregulate Nrf2/ARE Pathway.

doi: 10.3390/antiox11102055

Figure Lengend Snippet: Figure 5. Wine metabolites activate Nrf2/ARE pathway in basal conditions and in cells exposed to oxidative stress. (a,b) Effect of wine extract (WE) and sulforaphane (SFN) on Nrf2 nuclear translocation. THP-1 cells and cardiomyocytes were pre-incubated with WE, containing 800 µg/mL of total polyphenols and 10 µM SFN with or without the addition of tert-butyl hydroperoxide (TBHP, 100 µM). (c) Effect of WE on mRNA expression of Nrf2 target genes hemeoxygenase-1 (HO-1) and glutamate-cysteine ligase catalytic subunit (GCLC). (d) Average quantification of HO-1 and GCLC protein expression obtained by densitometric analysis and representative Western blot analyses for HO-1 and GCLC protein expression in cardiomyocytes. Data represent the mean +/−standard deviation of measurements performed in triplicate in three different experiments and; * p < 0.01 decrease vs. control; § p < 0.01 increase vs. control; ** p < 0.01 vs. TBHP.

Article Snippet: Human cardiomyocyte ventricular primary cells (Celprogen, Torrance, CA, USA) were cultured following the recommended manufacturer’s protocols in extracellular matrix precoated flasks and/or well plates with a ready-to-use complete growth medium (Celprogen, Torrance, CA, USA), as previously described [37].

Techniques: Translocation Assay, Incubation, Expressing, Western Blot, Standard Deviation, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Transcriptional Studies on Trypanosoma cruzi – Host Cell Interactions: A Complex Puzzle of Variables

doi: 10.3389/fcimb.2021.692134

Figure Lengend Snippet: Transcriptomic and functional results from different studies related to the host cell respiration response to T. cruzi infection.

Article Snippet: 4 , Dm28c/TcI , Vero , Primary human cardiomyocytes (CelProgen) , 10:1 , 2h , 0, 3, 6, 12hpi , Microarrays (Agilent) , Up-regulation of OXPHOS related genes ( ) .

Techniques: Functional Assay, Infection, In Vitro, In Vivo, Sequencing, Staining, Inhibition, Western Blot, Immunohistochemistry

Journal: Frontiers in Pharmacology

Article Title: Oleoylethanolamide as a New Therapeutic Strategy to Alleviate Doxorubicin-Induced Cardiotoxicity

doi: 10.3389/fphar.2022.863322

Figure Lengend Snippet: OEA inhibited DOX-induced oxidative stress and apoptosis in cardiomyocytes through TRPV1 pathway. HL-1 cells were treated with 0.1% DMSO, OEA (30 μM), GW6471 (5 μM), capsazepine (10 μM), nonivamide (10 μM), fenofibrate (100 μM) for 30 min. Cells were then treated with DOX (2 μM) for 24 h. Levels of (A) GST (B) TBARS and (C) cell viability was assessed in HL-1 cardiomyocytes. (D) TUNEL positive cells shown in (E) were counted. (E) TUNEL staining of HL-1 cardiomyocytes. Data are expressed as mean ± SEM, n = 4. ***, p < 0.001 vs. control. ##, p < 0.01; ###, p < 0.001 vs. DOX + vehicle. $$$, p < 0.001 vs. DOX + OEA.

Article Snippet: Adult derived primary human cardiomyocytes (Celprogen, Cat # 36044-15) were cultured in complete growth media (Celprogen, Cat #M36044-15S) in an incubator at 37°C with 5% CO 2 atmosphere.

Techniques: TUNEL Assay, Staining, Control

Journal: Frontiers in Pharmacology

Article Title: Oleoylethanolamide as a New Therapeutic Strategy to Alleviate Doxorubicin-Induced Cardiotoxicity

doi: 10.3389/fphar.2022.863322

Figure Lengend Snippet: OEA promotes TRPV1-mediated PI3K/ Akt signaling in cardiomyocytes. HL-1 cells were treated with 0.1% DMSO, OEA (30 μM), GW6471 (5 μM), capsazepine (10 μM) and nonivamide (10 μM) for 30 min. Cells were then treated with DOX (2 μM) for 24 h. PI3K, Akt, p-PI3K and p-Akt was determined using western blot analysis.

Article Snippet: Adult derived primary human cardiomyocytes (Celprogen, Cat # 36044-15) were cultured in complete growth media (Celprogen, Cat #M36044-15S) in an incubator at 37°C with 5% CO 2 atmosphere.

Techniques: Western Blot

Journal: Frontiers in Pharmacology

Article Title: Oleoylethanolamide as a New Therapeutic Strategy to Alleviate Doxorubicin-Induced Cardiotoxicity

doi: 10.3389/fphar.2022.863322

Figure Lengend Snippet: Inhibition of the PI3K-Akt signaling blocks the protective effects of OEA in cardiomyocytes. HL-1 cells were treated with 0.1% DMSO, OEA (30 μM) and LY294002 (10 μM) for 30 min. Cells were then treated with DOX (2 μM) for 24 h. Levels of (A) GST (B) TBARS and (C) cell viability was assessed in HL-1 cardiomyocytes. (D) TUNEL positive cells shown in (F) were counted. (E) Fluorescence quantification for caspase three in (G) were calculated. (F) TUNEL staining of HL-1 cardiomyocytes. (G) Immunofluorescence staining of caspase three in HL-1 cardiomyocytes. Data are expressed as mean ± SEM, n = 4. ***, p < 0.001 vs. control. #, p < 0.05; ##, p < 0.01; ###, p < 0.001 vs. DOX + vehicle. $, p < 0.05; $$, p < 0.01; $$$, p < 0.001 vs. DOX + OEA.

Article Snippet: Adult derived primary human cardiomyocytes (Celprogen, Cat # 36044-15) were cultured in complete growth media (Celprogen, Cat #M36044-15S) in an incubator at 37°C with 5% CO 2 atmosphere.

Techniques: Inhibition, TUNEL Assay, Fluorescence, Staining, Immunofluorescence, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Xenogeneic-Free System for Biomanufacturing of Cardiomyocyte Progeny From Human Pluripotent Stem Cells

doi: 10.3389/fbioe.2020.571425

Figure Lengend Snippet: Differentiation of hPSC-derived MoPS cells toward CMs: After 12–24 h of treatment with BMP4 (30 ng/ml) and Wnt3A (100 ng/ml), canonical Wnt signaling is inhibited with KY02111 (10 μM) from days 2–8. (A) Expression of early cardiac mesoderm (day 3) in hPSC-derived cells. (B) Representative plots of NKX2.5 expression in cells from H9 (21.8 ± 4.08%), IMR90-4 (11.61 ± 2.73%), and B12-3 (7.87 ± 3.8%) cells during transition through the cardiac mesoderm stage (see ). Results are shown as mean ± SD ( n ≥ 3). (C) Relative CM gene expression on day 16 of differentiation. (D) Flow cytometry analysis of differentiated H9 (91 ± 0.71%), IMR90-4 (81.31 ± 1.32%) and B12-3 (84.57 ± 5.12) hPSCs for TNNT2 expression. Black curve, isotype control. (E) Western blot of GATA4 (∼60 kDa) at the end of differentiation for hPSCs (H9:1-2, IMR90-4:3-4, B12-3 hPSCs: 5–6; directed differentiation: 1,3,5; no differentiation factor controls: 2,4,6; L: ladder). ACTB (∼40 kDa) is also shown.

Article Snippet: Samples were incubated at 4°C with primary antibodies for: cardiac troponin T (TNNT2; rabbit; ab45932; Abcam, Cambridge, MA), α-actinin (ACTN1; mouse; sc-15335) and GATA4 (rabbit; sc-9053; both from Santa Cruz Biotechnology, Dallas, TX).

Techniques: Derivative Assay, Expressing, Flow Cytometry, Western Blot

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Xenogeneic-Free System for Biomanufacturing of Cardiomyocyte Progeny From Human Pluripotent Stem Cells

doi: 10.3389/fbioe.2020.571425

Figure Lengend Snippet: Human PSC-derived CMs in planar culture. (A) Immunostaining of differentiated B12-3 cells for ACTN1. Bars: 50 μm. (B) Contractile activity of generated CMs in response to forskolin or isoproterenol. * p < 0.05, ** p < 0.01 vs. untreated cells. (C) Comparison of the number of live cells and viability between the XF protocol developed here (cell number: gray bars; viability: triangles/solid lines) and GW protocol (cell number: white bars; viability: squares/dashed lines) . Day 0 denotes the start of the differentiation. (D) Flow cytometry data depicting the fractions of TNNT2 + cells produced with the GW protocol (day 13) on vitronectin (top row) and Matrigel (bottom row).

Article Snippet: Samples were incubated at 4°C with primary antibodies for: cardiac troponin T (TNNT2; rabbit; ab45932; Abcam, Cambridge, MA), α-actinin (ACTN1; mouse; sc-15335) and GATA4 (rabbit; sc-9053; both from Santa Cruz Biotechnology, Dallas, TX).

Techniques: Derivative Assay, Immunostaining, Activity Assay, Generated, Flow Cytometry, Produced

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Xenogeneic-Free System for Biomanufacturing of Cardiomyocyte Progeny From Human Pluripotent Stem Cells

doi: 10.3389/fbioe.2020.571425

Figure Lengend Snippet: (A) Schematic of the hPSC differentiation process in stirred suspension culture. (B) Cell proliferation and viability of hPSCs expanded for 5 days and subsequently differentiated toward CMs. Both steps were carried out in spinner flasks. Solid lines represent live cell concentration and dashed lines represent viability. A representative run is shown. H9, gray curves; B12-3, red curves; IMR90-4, black curves. (C) Flow cytometry analysis of populations derived from the three hPSC lines on day 13 of differentiation for expression of: TNNT2 – H9: 92.32 ± 1.67%, IMR90-4: 89.13 ± 2.61%, B12-3: 86.69 ± 1.14%, and MYH1E – H9: 84.61 ± 2.59%, IMR90-4: 93.7 ± 3.34%, B12-3: 87.62 ± 3.49% (mean ± SD from n ≥ 3). Representative graphs are shown. Sample, red curve; isotype, black curve.

Article Snippet: Samples were incubated at 4°C with primary antibodies for: cardiac troponin T (TNNT2; rabbit; ab45932; Abcam, Cambridge, MA), α-actinin (ACTN1; mouse; sc-15335) and GATA4 (rabbit; sc-9053; both from Santa Cruz Biotechnology, Dallas, TX).

Techniques: Concentration Assay, Flow Cytometry, Derivative Assay, Expressing

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Xenogeneic-Free System for Biomanufacturing of Cardiomyocyte Progeny From Human Pluripotent Stem Cells

doi: 10.3389/fbioe.2020.571425

Figure Lengend Snippet: (A) Immunostaining of cells generated by the cardiogenic differentiation of hPSCs in spinner flasks. Representative micrographs are shown of ACTN1 (B12-3), TNNT2 (H9), and GATA4 (IMR90-4). DAPI staining of nuclear DNA is also shown. Bars: 50 μm. (B) Patch-clamp analysis of H9-derived CMs. A representative action-potential graph is shown.

Article Snippet: Samples were incubated at 4°C with primary antibodies for: cardiac troponin T (TNNT2; rabbit; ab45932; Abcam, Cambridge, MA), α-actinin (ACTN1; mouse; sc-15335) and GATA4 (rabbit; sc-9053; both from Santa Cruz Biotechnology, Dallas, TX).

Techniques: Immunostaining, Generated, Staining, Patch Clamp, Derivative Assay