Journal: International Journal of Molecular Sciences

Article Title: An Evaluation of Human Induced Pluripotent Stem Cells to Test for Cardiac Developmental Toxicity

doi: 10.3390/ijms22158114

Figure Lengend Snippet: hiPSCs consistently and robustly differentiated into cardiomyocytes. Differentiated cardiomyocytes were assessed for cardiomyocyte-specific markers and gene expression. ( A ) Schematic protocol for cardiomyocyte differentiation and embryotoxicity screen. ( B ) Immunocytochemistry stains of differentiated cultures confirmed cardiomyocyte identity via myosin heavy chain (MHC) and Troponin I (Trop I). ( C ) Differentiated cardiomyocytes expressed cardiac-specific genes TBX5 and MEF2c as measured by qPCR, n = 3 biological replicates ± SD, * p < 0.05, Student’s t -test. hiPSC, human induced pluripotent stem cell; MHC, myosin heavy chain; Trop I, Troponin I.

Article Snippet: The hiPSC cell line RIV9 was obtained from the Stem Cell Core at the University of California, Riverside. hiPSCs were seeded on Matrigel (Corning, Glendale, AZ, USA)-coated tissue culture plates and maintained in a pluripotent state in mTeSR ® medium (Stem Cell Technologies, Seattle, WA, USA).

Techniques: Gene Expression, Immunocytochemistry

Journal: International Journal of Molecular Sciences

Article Title: An Evaluation of Human Induced Pluripotent Stem Cells to Test for Cardiac Developmental Toxicity

doi: 10.3390/ijms22158114

Figure Lengend Snippet: Presence of contractile cardiomyocyte clusters declines with treatment with embryotoxicants 5-FU and at RA. hiPSCs were treated with different concentrations of 5-FU, at RA, or PenG and photographed. hiPSC, human induced pluripotent stem cell; 5-FU, 5-fluorouracil; at RA, all-trans retinoic acid; PenG, penicillin G.

Article Snippet: The hiPSC cell line RIV9 was obtained from the Stem Cell Core at the University of California, Riverside. hiPSCs were seeded on Matrigel (Corning, Glendale, AZ, USA)-coated tissue culture plates and maintained in a pluripotent state in mTeSR ® medium (Stem Cell Technologies, Seattle, WA, USA).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: An Evaluation of Human Induced Pluripotent Stem Cells to Test for Cardiac Developmental Toxicity

doi: 10.3390/ijms22158114

Figure Lengend Snippet: Treatment with embryotoxicants 5-FU and at RA impeded cardiomyocyte differentiation. hiPSCs were treated with different concentrations of 5-FU, at RA, or PenG and evaluated for differentiation inhibition by visually scoring the number of actively contracting cardiomyocyte clusters. Each data point represents the mean of three independent experiments ± SD. *: p < 0.05 = the lowest concentration significantly below the untreated control group as determined by One-Way ANOVA. hiPSC, human induced pluripotent stem cell; 5-FU, 5-fluorouracil; at RA, all-trans retinoic acid; PenG, penicillin G.

Article Snippet: The hiPSC cell line RIV9 was obtained from the Stem Cell Core at the University of California, Riverside. hiPSCs were seeded on Matrigel (Corning, Glendale, AZ, USA)-coated tissue culture plates and maintained in a pluripotent state in mTeSR ® medium (Stem Cell Technologies, Seattle, WA, USA).

Techniques: Inhibition, Concentration Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: An Evaluation of Human Induced Pluripotent Stem Cells to Test for Cardiac Developmental Toxicity

doi: 10.3390/ijms22158114

Figure Lengend Snippet: Comparison of mESC- and hiPSC-EST IC 50 and ID 50 values and embryotoxicity classifications.

Article Snippet: The hiPSC cell line RIV9 was obtained from the Stem Cell Core at the University of California, Riverside. hiPSCs were seeded on Matrigel (Corning, Glendale, AZ, USA)-coated tissue culture plates and maintained in a pluripotent state in mTeSR ® medium (Stem Cell Technologies, Seattle, WA, USA).

Techniques: Comparison

Journal: International Journal of Molecular Sciences

Article Title: An Evaluation of Human Induced Pluripotent Stem Cells to Test for Cardiac Developmental Toxicity

doi: 10.3390/ijms22158114

Figure Lengend Snippet: 5-FU and at RA treatment reduced hiPSC-cardiomyocyte and hFF viability in a dose-dependent manner as assessed via MTT assay. ( A ) hiPSC viability screens for 5-FU, at RA, and PenG, n = 3 biological replicates ± SD. *: p < 0.05 = the lowest concentration significantly below the untreated hiPSC control group as determined by One-Way ANOVA. ( B ) hFF viability screens for 5-FU, at RA, and PenG, n = 3 biological replicates ± SD. *: p < 0.05 = the lowest concentration significantly below the untreated hFF control group as determined by One-Way ANOVA. hiPSC, human induced pluripotent stem cell; MTT, mitochondrial dehydrogenase activity assay; 5-FU, 5-fluorouracil; at RA, all-trans retinoic acid; PenG, penicillin G; hFF, human foreskin fibroblast.

Article Snippet: The hiPSC cell line RIV9 was obtained from the Stem Cell Core at the University of California, Riverside. hiPSCs were seeded on Matrigel (Corning, Glendale, AZ, USA)-coated tissue culture plates and maintained in a pluripotent state in mTeSR ® medium (Stem Cell Technologies, Seattle, WA, USA).

Techniques: MTT Assay, Concentration Assay, Control, MTT Mitochondrial, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: An Evaluation of Human Induced Pluripotent Stem Cells to Test for Cardiac Developmental Toxicity

doi: 10.3390/ijms22158114

Figure Lengend Snippet: Treatment with embryotoxicants 5-FU and at RA impeded cardiomyocyte differentiation as measured by day 10 TBX5 gene expression in hiPSCs. hiPSCs were treated with different concentrations of 5-FU, at RA, or PenG and evaluated for TBX5 or MEF2c expression via qPCR. Data points represent means of three independent experiments ± SD. Inhibition of differentiation (ID 50 ) was determined from dose–response curves as a 50% reduction in gene expression in the control. hiPSC, human induced pluripotent stem cell; 5-FU, 5-fluorouracil; at RA, all-trans retinoic acid; PenG, penicillin G. *: p < 0.05 = the lowest concentration significantly below the untreated hiPSC control group as determined by One-Way ANOVA.

Article Snippet: The hiPSC cell line RIV9 was obtained from the Stem Cell Core at the University of California, Riverside. hiPSCs were seeded on Matrigel (Corning, Glendale, AZ, USA)-coated tissue culture plates and maintained in a pluripotent state in mTeSR ® medium (Stem Cell Technologies, Seattle, WA, USA).

Techniques: Gene Expression, Expressing, Inhibition, Control, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: An Evaluation of Human Induced Pluripotent Stem Cells to Test for Cardiac Developmental Toxicity

doi: 10.3390/ijms22158114

Figure Lengend Snippet: List of IC 50 and ID 50 values and embryotoxicity classifications determined from concentration-response curves for contractile and d10 qPCR assay endpoints. hiPSC, human induced pluripotent stem cell; hFF, human foreskin fibroblast.

Article Snippet: The hiPSC cell line RIV9 was obtained from the Stem Cell Core at the University of California, Riverside. hiPSCs were seeded on Matrigel (Corning, Glendale, AZ, USA)-coated tissue culture plates and maintained in a pluripotent state in mTeSR ® medium (Stem Cell Technologies, Seattle, WA, USA).

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: An Evaluation of Human Induced Pluripotent Stem Cells to Test for Cardiac Developmental Toxicity

doi: 10.3390/ijms22158114

Figure Lengend Snippet: Effects of cigarette smoke and Snus smokeless tobacco on developing cardiomyocytes. ( A ) Contractile and viability screens for hiPSCs exposed to MS cigarette smoke using contractile and MTT assay, n = 3 ± SD. ( B ) Contractile and viability screens for hiPSCs exposed to Snus smokeless tobacco using contractile and MTT assay, n = 3 ± SD. ( C ) Viability screen for hFFs exposed to MS cigarette smoke using MTT assay, n = 3 ± SD. ( D ) Viability screen for hFFs exposed to Snus smokeless tobacco using MTT assay, n = 3 ± SD. hiPSC, human induced pluripotent stem cell; MTT, mitochondrial dehydrogenase activity assay; MS; mainstream; hFF, human foreskin fibroblast. *: p < 0.05 = the lowest concentration significantly below the untreated hiPSC control group as determined by One-Way ANOVA.

Article Snippet: The hiPSC cell line RIV9 was obtained from the Stem Cell Core at the University of California, Riverside. hiPSCs were seeded on Matrigel (Corning, Glendale, AZ, USA)-coated tissue culture plates and maintained in a pluripotent state in mTeSR ® medium (Stem Cell Technologies, Seattle, WA, USA).

Techniques: MTT Assay, MTT Mitochondrial, Activity Assay, Concentration Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: An Evaluation of Human Induced Pluripotent Stem Cells to Test for Cardiac Developmental Toxicity

doi: 10.3390/ijms22158114

Figure Lengend Snippet: List of IC 50 and ID 50 values determined from concentration-response curves for mainstream cigarette smoke and Snus smokeless tobacco. hiPSC, human induced pluripotent stem cell; hFF, human foreskin fibroblast.

Article Snippet: The hiPSC cell line RIV9 was obtained from the Stem Cell Core at the University of California, Riverside. hiPSCs were seeded on Matrigel (Corning, Glendale, AZ, USA)-coated tissue culture plates and maintained in a pluripotent state in mTeSR ® medium (Stem Cell Technologies, Seattle, WA, USA).

Techniques: Concentration Assay

Journal: Cells

Article Title: Neurodegeneration Induced by Anti-IgLON5 Antibodies Studied in Induced Pluripotent Stem Cell-Derived Human Neurons

doi: 10.3390/cells10040837

Figure Lengend Snippet: Graphical illustration of the experimental setup. Short-term autoantibody exposure was performed on human Neuronal Stem cells (hNSC) and the experimental setup is shown in panel ( A ). Long-term autoantibody exposure was performed on human-induced Pluripotent Stem Cell (hiPSC)-derived neurons and the experimental setup is shown in panel ( B ). Black arrows and text specify differentiation procedures. Colored arrows and text boxes specify the time points at which different analyses were performed. LDH: lactate dehydrogenase, MEA: multi electrode array.

Article Snippet: For long-term exposure experiments, a predifferentiated human-induced pluripotent stem cell line (hiPSC, XCell Science, Novato, CA, USA) was propagated in Geltrex (Gibco, Thermo Fisher, Waltham, MA, USA)-coated 6-well plates [ ].

Techniques: Derivative Assay

Journal: Cells

Article Title: Neurodegeneration Induced by Anti-IgLON5 Antibodies Studied in Induced Pluripotent Stem Cell-Derived Human Neurons

doi: 10.3390/cells10040837

Figure Lengend Snippet: Anti-IgLON5 antibodies reduce cell surface IgLON5 clusters, synaptic proteins, and neuronal activity. Indirect immunofluorescent staining of human induced pluripotent stem cell (hiPSC) cultures treated with control IgG or anti-IgLON5 IgG ( A ). Quantification of IgLON5-positive clusters in hiPSC cultures revealed a significant reduction after 21 and 35 days of antibody exposure ( B ) ( n = 9). Immunostaining for the synaptic proteins synaptophysin and PSD95 revealed a decrease in synaptophysin clusters after 21 days of exposure and in both synaptophysin and PSD95 clusters after 35 days of exposure ( C , D ) ( n = 6). Multi electrode array (MEA) analysis of hiPSC cultures showed the expected increase in neuronal activity as they matured, however, the spike rate of neurons treated with anti-IgLON5 IgG was significantly lower than that of untreated neurons ( E ) ( n = 5). Example of a MEA data sample is shown in ( F ) (top). When the signal crosses the threshold (horizontal lines) a spike is registered. Registered spikes from a single electrode in a well with an untreated culture (F, middle) and an anti-IgLON5 IgG-treated culture (F, bottom) during a 15 s period. Statistical analysis: two-tailed unpaired t -test or Two-way ANOVA followed by Sidak’s multiple comparisons test (*: p < 0.05, ***: p < 0.001, ****: p < 0.0001). Scale bar length: 5 μm.

Article Snippet: For long-term exposure experiments, a predifferentiated human-induced pluripotent stem cell line (hiPSC, XCell Science, Novato, CA, USA) was propagated in Geltrex (Gibco, Thermo Fisher, Waltham, MA, USA)-coated 6-well plates [ ].

Techniques: Activity Assay, Staining, Control, Immunostaining, Two Tailed Test

Journal: Cells

Article Title: Neurodegeneration Induced by Anti-IgLON5 Antibodies Studied in Induced Pluripotent Stem Cell-Derived Human Neurons

doi: 10.3390/cells10040837

Figure Lengend Snippet: Anti-IgLON5 antibodies increase cell death in induced pluripotent stem cell (hiPSC)-derived neural cultures. Cultures treated with anti-IgLON5 IgG showed an increase in phosphorylated-tau (p-tau) (T205) expression after 7 and 21 days of exposure ( A – C ) ( n = 7–8). Quantification of nuclei with unhealthy morphology revealed an increased percentage of dying cells after 21 and 35 days of exposure ( D ) ( n = 7–18). A lactate dehydrogenase (LDH)-assay (a marker of necrosis) showed increased LDH release to the culture medium of anti-IgLON5 IgG exposed cultures after 30 days ( E ) ( n = 8). Statistical analysis: two-way ANOVA followed by Sidak’s multiple comparisons test. (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). Scale bars: 50 μm. β-tub III: β-tubulin III.

Article Snippet: For long-term exposure experiments, a predifferentiated human-induced pluripotent stem cell line (hiPSC, XCell Science, Novato, CA, USA) was propagated in Geltrex (Gibco, Thermo Fisher, Waltham, MA, USA)-coated 6-well plates [ ].

Techniques: Derivative Assay, Expressing, Lactate Dehydrogenase Assay, Marker