Journal: International Journal of Molecular Sciences

Article Title: A Novel In Vitro Assay Using Human iPSC-Derived Sensory Neurons to Evaluate the Effects of External Chemicals on Neuronal Morphology: Possible Implications in the Prediction of Abnormal Skin Sensation

doi: 10.3390/ijms221910525

Figure Lengend Snippet: Effects of direct action of MP on nerve fiber elongation of hiPSC-SNs. After culturing for 3 days, hiPSC-SNs were incubated with medium containing MP for 24 h and stained with NeuO. ( a ) Representative images with nerve fibers outlined in red were obtained using IN Cell Analyzer 2200. The red lines indicate NeuO-stained nerve fibers. ( b ) Nerve fiber length per cell versus the control after incubation with various doses of MP. Each value represents the mean ± standard deviation of 4–25 wells. ** p < 0.005, *** p < 0.001, vs. the control (Steel’s test). MP: methylparaben.

Article Snippet: After 2 days of culture, the medium was replaced with ReproNeuro MQ medium containing one of the following: 0.0004% (0.03 mM), 0.002% (0.13 mM), 0.01% (0.66 mM), or 0.05% (3.3 mM) methylparaben (132-02635; FUJIFILM Wako, Osaka, Japan); 0.01% (0.7 mM), 0.04% (3 mM), or 0.5% (36 mM) phenoxyethanol (169-12072; FUJIFILM Wako, Osaka, Japan); or 0.5, 3, or 18 μM benzo[a]pyrene (BaP, 50-32-8; FUJIFILM Wako, Osaka, Japan) with 0.05% dimethyl sulphoxide (DMSO, 94563; Merck KGaA, Darmstadt, Germany); or 0.05% DMSO (as a control) and incubated for 24 h at 37 °C under 5% CO 2 .

Techniques: Incubation, Staining, Control, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: A Novel In Vitro Assay Using Human iPSC-Derived Sensory Neurons to Evaluate the Effects of External Chemicals on Neuronal Morphology: Possible Implications in the Prediction of Abnormal Skin Sensation

doi: 10.3390/ijms221910525

Figure Lengend Snippet: Effects of direct action of PE on nerve fiber elongation of hiPSC-SNs. After culturing for 3 days, hiPSC-SNs were incubated with medium containing methylparaben for 24 h and stained with NeuO. ( a ) Representative images with nerve fibers outlined in red were obtained using IN Cell Analyzer 2200. The red lines indicate NeuO-stained nerve fibers. ( b ) Nerve fiber length per cell versus the control after incubation with various doses of PE. Each value represents the mean ± standard deviation of 7–25 wells. *** p < 0.001, vs. the control (Steel’s test). PE: phenoxyethanol.

Article Snippet: After 2 days of culture, the medium was replaced with ReproNeuro MQ medium containing one of the following: 0.0004% (0.03 mM), 0.002% (0.13 mM), 0.01% (0.66 mM), or 0.05% (3.3 mM) methylparaben (132-02635; FUJIFILM Wako, Osaka, Japan); 0.01% (0.7 mM), 0.04% (3 mM), or 0.5% (36 mM) phenoxyethanol (169-12072; FUJIFILM Wako, Osaka, Japan); or 0.5, 3, or 18 μM benzo[a]pyrene (BaP, 50-32-8; FUJIFILM Wako, Osaka, Japan) with 0.05% dimethyl sulphoxide (DMSO, 94563; Merck KGaA, Darmstadt, Germany); or 0.05% DMSO (as a control) and incubated for 24 h at 37 °C under 5% CO 2 .

Techniques: Incubation, Staining, Control, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: A Novel In Vitro Assay Using Human iPSC-Derived Sensory Neurons to Evaluate the Effects of External Chemicals on Neuronal Morphology: Possible Implications in the Prediction of Abnormal Skin Sensation

doi: 10.3390/ijms221910525

Figure Lengend Snippet: Effects of MP treatment on bleb formation and NMNAT2 expression in nerve fibers. After culturing for 3 days, the effects of the direct interaction between hiPSC-SN and MP were assessed. ( a ) Morphological changes in nerve fibers 24 h after treatment with MP. The arrows indicate blebs. ( b ) The number of blebs at various concentrations of MP. *** p < 0.001 vs. absence of MP (Dunnett’s test). ( c ) Double labelling of NMNAT2 (red) and βIII-tubulin (green) in nerve fibers at various concentrations of MP. ( d ) NMNAT2-immunoreactive (ir) nerve fiber length normalised to βIII-tubulin-ir nerve fiber length in identical areas at various concentrations of MP. ** p < 0.005 vs. absence of MP (Williams’ test). Each value represents the mean ± standard deviation of more than three wells. MP: methylparaben; NMNAT2: nicotinamide mononucleotide adenylyltransferase 2.

Article Snippet: After 2 days of culture, the medium was replaced with ReproNeuro MQ medium containing one of the following: 0.0004% (0.03 mM), 0.002% (0.13 mM), 0.01% (0.66 mM), or 0.05% (3.3 mM) methylparaben (132-02635; FUJIFILM Wako, Osaka, Japan); 0.01% (0.7 mM), 0.04% (3 mM), or 0.5% (36 mM) phenoxyethanol (169-12072; FUJIFILM Wako, Osaka, Japan); or 0.5, 3, or 18 μM benzo[a]pyrene (BaP, 50-32-8; FUJIFILM Wako, Osaka, Japan) with 0.05% dimethyl sulphoxide (DMSO, 94563; Merck KGaA, Darmstadt, Germany); or 0.05% DMSO (as a control) and incubated for 24 h at 37 °C under 5% CO 2 .

Techniques: Expressing, Standard Deviation