Journal: Journal of Cancer

Article Title: Talin1 Mediates Tumor-Nerve Interactions in Prostate and Breast Cancer Cells

doi: 10.7150/jca.127292

Figure Lengend Snippet: Human Neural Progenitor Cells (NPCs) display increased neurite outgrowth when cultured with exosomes and CM from MCF-7 Snail cells compared to MCF-7 Neo. MCF-7 Neo or MCF-7 Snail was cultured in exosome-depleted medium (Exo-D medium) for 3 days, and conditioned medium (CM) was collected, or exosomes were isolated from the CM. These were subsequently cultured with NPCs for 2 days. (A) Images of NPC after incubation in NPC regular medium, Exo-D medium, exosomes or CM with or without Talin1 inhibitor mH4 were captured with a Nikon Eclipse TE2000-S Inverted Fluorescence Microscope. (B) The percentage of NPCs with neurite outgrowth and average neuron length was measured utilizing Image J Fiji Just software in NPCs cultured with NPC medium, Exo-D medium, and Exo-D medium plus NGF-beta as a positive control. The percentage of NPCs with neurite outgrowth and average neuron length of NPCs cultured with (C) exosomes isolated from MCF7-Neo and MCF7-Snail CM or (D) CM from MCF7-Neo and MCF7-Snail was analyzed using ImageJ Fiji Just software. Statistical analyses were done using GraphPad Prism software (one-way ANOVA, Šídák's multiple comparisons test, ****p< 0.0001, ***p< 0.001, **p < 0.01, *p < 0.05). Bars represent the SD of the mean. Results are representative of 3 experiments performed independently.

Article Snippet: Neural Progenitor Cells (ACS-5004), NPC Growth Medium (ATCC ACS-3003), DMEM: F12 (ATCC 30-2006) and Cell Basement Membrane (ACS-3035) were purchased from American Tissue Cell Culture (ATCC), Manassas, VA. Talin1 inhibitor, mH4 and F3 protease inhibitor control, was obtained from Dr. Kensei Tsuzaka, Kaytee Bio, Co. & Ltd., Chiba, Japan. mH4 was dissolved in DMSO to a stock solution of 14 mg/ml.

Techniques: Cell Culture, Isolation, Incubation, Fluorescence, Microscopy, Software, Positive Control

Journal: Journal of Cancer

Article Title: Talin1 Mediates Tumor-Nerve Interactions in Prostate and Breast Cancer Cells

doi: 10.7150/jca.127292

Figure Lengend Snippet: Human neural progenitor cells (NPCs) display reduced neurite outgrowth with exosomes and CM from C4-2 Snail knockdown (C4-2 E8) cells compared to C4-2 Non-silencing (NS) cells. C4-2 NS or C4-2 E8 was cultured in exosome-depleted medium (Exo-D medium) for 3 days, and conditioned medium (CM) was collected, or exosomes were isolated from the CM. These were subsequently cultured with NPCs for 2 days. (A) Images of NPC after incubation in NPC regular medium, Exo-D medium, exosomes, or CM with or without Talin1 inhibitor mH4 were captured with a Nikon Eclipse TE2000-S Inverted Fluorescence Microscope. The percentage of NPCs with neurite outgrowth and average neuron length of NPCs cultured with (B) exosomes isolated from MCF7-Neo and MCF7-Snail CM or (C) CM from MCF7-Neo and MCF7-Snail was analyzed using ImageJ Fiji Just software. Statistical analyses were done using GraphPad Prism software (one-way ANOVA, Šídák's multiple comparisons test, ****p< 0.0001, **p < 0.01, *p < 0.05). Bars represent the SD of the mean. Results are representative of 3 experiments performed independently.

Article Snippet: Neural Progenitor Cells (ACS-5004), NPC Growth Medium (ATCC ACS-3003), DMEM: F12 (ATCC 30-2006) and Cell Basement Membrane (ACS-3035) were purchased from American Tissue Cell Culture (ATCC), Manassas, VA. Talin1 inhibitor, mH4 and F3 protease inhibitor control, was obtained from Dr. Kensei Tsuzaka, Kaytee Bio, Co. & Ltd., Chiba, Japan. mH4 was dissolved in DMSO to a stock solution of 14 mg/ml.

Techniques: Knockdown, Cell Culture, Isolation, Incubation, Fluorescence, Microscopy, Software

Journal: Cancers

Article Title: Glioblastoma Cells Induce Neuron Loss In Vivo and In Vitro

doi: 10.3390/cancers17172817

Figure Lengend Snippet: Establishment of the GSC–neuron coculture. ( a ) Neuron differentiation: iPSC-derived NPCs were induced to differentiate into neurons for three weeks and were characterized for the neuronal markers Tuj1 (green) and NeuN (red). Cell nuclei were stained with DAPI (blue). ( b ) Establishment of the GSC–neuron coculture: NeuN + neurons were used for the cocultures. GFP + GSCs (NSC11 and NSC20) were added directly to the neurons in the GSC media for 48 h to establish the coculture. The figure shows representative images of the GSCs (GFP: green)–neurons (NeuN: red) in the direct coculture. The images were captured with a confocal microscope using a 40× lens, and images of the neurons are presented as orthogonal projections.

Article Snippet: iPSC-derived NPCs (Catalog #ACS5004) were procured from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Derivative Assay, Staining, Microscopy

Journal: Journal of Nanobiotechnology

Article Title: Development of NIR photocleavable nanoparticles with BDNF for vestibular neuron regeneration

doi: 10.1186/s12951-025-03298-x

Figure Lengend Snippet: UV (254 nm) vs. laser (808 nm) cytotoxicity assay in human neural progenitor cells (hNPCs). ( a ) hNPCs were grown in culture for 3 days prior to UV or laser treatment, and viability assays were performed 24 h after. Immunofluorescence images show the expression of NPC markers SOX1 and Nestin. ( b ) UV irradiation at energy densities 30, 60, & 90 J/cm 2 significantly decreased the viability of hNPCs after 24 h in a culture determined via CCK-8 assay. ( c ) Laser irradiation at energy densities 30, 60, & 90 J/cm 2 did not affect hNPCs viability. The statistical significance of the data was analyzed using one-way ANOVA with Tukey’s multiple comparisons test for each group (**** P < 0.0001)

Article Snippet: hNPCs (ACS-5004; ATCC, Manassas, VA, USA) were used to determine the cytotoxicity of PCNs in culture.

Techniques: Cytotoxicity Assay, Immunofluorescence, Expressing, Irradiation, CCK-8 Assay

Journal: Journal of Nanobiotechnology

Article Title: Development of NIR photocleavable nanoparticles with BDNF for vestibular neuron regeneration

doi: 10.1186/s12951-025-03298-x

Figure Lengend Snippet: Photocleavable nanoparticle (PCN) cytotoxicity assay in human neural progenitor cells (hNPCs). ( a ) hNPCs were grown in culture for 3 days prior to PCN or laser treatment, and viability assays were performed after 24 h. ( b ) Concentrations of PCN up to 900 µg/mL exhibited no cytotoxic effect on hNPCs. ( c ) Laser irradiation in combination with PCNs at energy densities of 30, 60, and 90 J/cm 2 , did not affect hNPC viability after 24 h in culture, as determined by the CCK-8 assay. ( d ) Immunofluorescence images show the presence of PCNs in hNPCs, confirming that hNPCs remained viable after treatment and laser irradiation. Groups were compared using one-way ANOVA with Tukey’s multiple comparisons test

Article Snippet: hNPCs (ACS-5004; ATCC, Manassas, VA, USA) were used to determine the cytotoxicity of PCNs in culture.

Techniques: Cytotoxicity Assay, Irradiation, CCK-8 Assay, Immunofluorescence

Journal: Journal of nanobiotechnology

Article Title: Development of NIR photocleavable nanoparticles with BDNF for vestibular neuron regeneration.

doi: 10.1186/s12951-025-03298-x

Figure Lengend Snippet: Fig. 1 UV (254 nm) vs. laser (808 nm) cytotoxicity assay in human neural progenitor cells (hNPCs). (a) hNPCs were grown in culture for 3 days prior to UV or laser treatment, and viability assays were performed 24 h after. Immunofluorescence images show the expression of NPC markers SOX1 and Nestin. (b) UV irradiation at energy densities 30, 60, & 90 J/cm2 significantly decreased the viability of hNPCs after 24 h in a culture determined via CCK-8 assay. (c) Laser irradiation at energy densities 30, 60, & 90 J/cm2 did not affect hNPCs viability. The statistical significance of the data was analyzed using one-way ANOVA with Tukey’s multiple comparisons test for each group (**** P < 0.0001)

Article Snippet: Evaluation of cytotoxic effects of PCNs and laser treatment on hNPCs hNPCs (ACS-5004; ATCC, Manassas, VA, USA) were used to determine the cytotoxicity of PCNs in culture.

Techniques: Cytotoxicity Assay, Immunofluorescence, Expressing, Irradiation, CCK-8 Assay

Journal: Journal of nanobiotechnology

Article Title: Development of NIR photocleavable nanoparticles with BDNF for vestibular neuron regeneration.

doi: 10.1186/s12951-025-03298-x

Figure Lengend Snippet: Fig. 5 Photocleavable nanoparticle (PCN) cytotoxicity assay in human neural progenitor cells (hNPCs). (a) hNPCs were grown in culture for 3 days prior to PCN or laser treatment, and viability assays were performed after 24 h. (b) Concentrations of PCN up to 900 µg/mL exhibited no cytotoxic effect on hNPCs. (c) Laser irradiation in combination with PCNs at energy densities of 30, 60, and 90 J/cm2, did not affect hNPC viability after 24 h in culture, as determined by the CCK-8 assay. (d) Immunofluorescence images show the presence of PCNs in hNPCs, confirming that hNPCs remained viable after treatment and laser irradiation. Groups were compared using one-way ANOVA with Tukey’s multiple comparisons test

Article Snippet: Evaluation of cytotoxic effects of PCNs and laser treatment on hNPCs hNPCs (ACS-5004; ATCC, Manassas, VA, USA) were used to determine the cytotoxicity of PCNs in culture.

Techniques: Cytotoxicity Assay, Irradiation, CCK-8 Assay, Immunofluorescence

Journal: Journal of nanobiotechnology

Article Title: Development of NIR photocleavable nanoparticles with BDNF for vestibular neuron regeneration.

doi: 10.1186/s12951-025-03298-x

Figure Lengend Snippet: Fig. 6 Phototriggered release of brain-derived neurotrophic factor (BDNF) from photocleavable nanoparticles (PCNs) and its effects on the growth and maturation of human neural progenitor cells (hNPCs). (a) Amounts (%) of BDNF released in hNPCs at different energy densities (0, 30, 60, and 90 J/cm2) after laser irradiation. Irradiation at 60 J/cm2 (89.15%) showed BDNF release that was higher relative to 30 J/cm2 (69.01%) but not significantly higher than 90 J/cm2 (90.76%). (b) hNPCs were grown in culture for 3 days prior to PCN or laser treatment, and the medium was changed to neuronal differentiation medium (NDM) containing no BDNF. Maturation effects were determined after 7 days in NDM (D10). (c) The neural progenitor cell marker Nestin had significantly higher mean fluorescence intensity relative to PCN only and PCN + 90 J/cm2, as assessed by immunohistochemical staining (d). (e) The cor responding immunofluorescence images also observed a higher mean fluorescence intensity with PCN + 60 J/cm2 for the mature neuronal cell marker NeuN (f). Groups were compared using one-way ANOVA with Tukey’s multiple comparisons test (**** P < 0.0001)

Article Snippet: Evaluation of cytotoxic effects of PCNs and laser treatment on hNPCs hNPCs (ACS-5004; ATCC, Manassas, VA, USA) were used to determine the cytotoxicity of PCNs in culture.

Techniques: Derivative Assay, Irradiation, Marker, Fluorescence, Immunohistochemical staining, Staining, Immunofluorescence