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Representative confocal images of fluorescent PS nanoplastic internalization after the indicated time in: ( A ) primary bone marrow CD34+ sorted hematopoietic immature cells or ( B ) <t>epithelial</t> or fibroblast cells isolated from primary mammary gland obtained from healthy donors or ( C ) MCF10A-derived models representing non-transformed mammary stem cells (MCF10A-CT), early (MC26) and more transformed (M1B26) cells as previously described . Nuclei are stained in blue, actin filaments in red, PS in green. ( D ) Confocal images of PS and PET nanoplastic internalization in MCF10A after 24 h compared to non-exposed controls. Nuclei are stained blue actin filaments red, PS green, PET orange. ( E ) Schematic diagram of the nanoplastic (NPLs) long-term exposure approach and analysis of their impact. ( F ) Quantification of the sphere-forming ability of untreated (CT) MCF10A cells or following 20 weeks of exposure to PS or PET nanoplastics (NPLs), n=5. ( G ) Quantification of total stem cell-derived progenitors by E-CFC assay , data represent the total number of colonies per 250 seeded cells, n=5. ( F , G , ) Each individual datum is represented. One way ANOVA is indicated on the graph by the P values. ( H ) Representation of the mean proportion of myoepithelial, mixed and luminal colonies quantified by E-CFC assay (n = 5). Data are expressed as the percentage of each subtype with a total number of scored colonies representing 100%. ( I ) Confocal images of acini. Nuclei are stained in blue, actin filaments in red, white arrows point to acini with a disorganized structure. Measurement of acini ( J ) solidity or ( K ) Ferret diameter ratio was quantified by image analysis using Fiji imaging software, n=109 to 131 individual acini. ( J , K ) Each individual data is represented. Mann-Whitney test is indicated on the graph by the P values.
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Representative confocal images of fluorescent PS nanoplastic internalization after the indicated time in: ( A ) primary bone marrow CD34+ sorted hematopoietic immature cells or ( B ) <t>epithelial</t> or fibroblast cells isolated from primary mammary gland obtained from healthy donors or ( C ) MCF10A-derived models representing non-transformed mammary stem cells (MCF10A-CT), early (MC26) and more transformed (M1B26) cells as previously described . Nuclei are stained in blue, actin filaments in red, PS in green. ( D ) Confocal images of PS and PET nanoplastic internalization in MCF10A after 24 h compared to non-exposed controls. Nuclei are stained blue actin filaments red, PS green, PET orange. ( E ) Schematic diagram of the nanoplastic (NPLs) long-term exposure approach and analysis of their impact. ( F ) Quantification of the sphere-forming ability of untreated (CT) MCF10A cells or following 20 weeks of exposure to PS or PET nanoplastics (NPLs), n=5. ( G ) Quantification of total stem cell-derived progenitors by E-CFC assay , data represent the total number of colonies per 250 seeded cells, n=5. ( F , G , ) Each individual datum is represented. One way ANOVA is indicated on the graph by the P values. ( H ) Representation of the mean proportion of myoepithelial, mixed and luminal colonies quantified by E-CFC assay (n = 5). Data are expressed as the percentage of each subtype with a total number of scored colonies representing 100%. ( I ) Confocal images of acini. Nuclei are stained in blue, actin filaments in red, white arrows point to acini with a disorganized structure. Measurement of acini ( J ) solidity or ( K ) Ferret diameter ratio was quantified by image analysis using Fiji imaging software, n=109 to 131 individual acini. ( J , K ) Each individual data is represented. Mann-Whitney test is indicated on the graph by the P values.
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Representative confocal images of fluorescent PS nanoplastic internalization after the indicated time in: ( A ) primary bone marrow CD34+ sorted hematopoietic immature cells or ( B ) <t>epithelial</t> or fibroblast cells isolated from primary mammary gland obtained from healthy donors or ( C ) MCF10A-derived models representing non-transformed mammary stem cells (MCF10A-CT), early (MC26) and more transformed (M1B26) cells as previously described . Nuclei are stained in blue, actin filaments in red, PS in green. ( D ) Confocal images of PS and PET nanoplastic internalization in MCF10A after 24 h compared to non-exposed controls. Nuclei are stained blue actin filaments red, PS green, PET orange. ( E ) Schematic diagram of the nanoplastic (NPLs) long-term exposure approach and analysis of their impact. ( F ) Quantification of the sphere-forming ability of untreated (CT) MCF10A cells or following 20 weeks of exposure to PS or PET nanoplastics (NPLs), n=5. ( G ) Quantification of total stem cell-derived progenitors by E-CFC assay , data represent the total number of colonies per 250 seeded cells, n=5. ( F , G , ) Each individual datum is represented. One way ANOVA is indicated on the graph by the P values. ( H ) Representation of the mean proportion of myoepithelial, mixed and luminal colonies quantified by E-CFC assay (n = 5). Data are expressed as the percentage of each subtype with a total number of scored colonies representing 100%. ( I ) Confocal images of acini. Nuclei are stained in blue, actin filaments in red, white arrows point to acini with a disorganized structure. Measurement of acini ( J ) solidity or ( K ) Ferret diameter ratio was quantified by image analysis using Fiji imaging software, n=109 to 131 individual acini. ( J , K ) Each individual data is represented. Mann-Whitney test is indicated on the graph by the P values.
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Representative confocal images of fluorescent PS nanoplastic internalization after the indicated time in: ( A ) primary bone marrow CD34+ sorted hematopoietic immature cells or ( B ) <t>epithelial</t> or fibroblast cells isolated from primary mammary gland obtained from healthy donors or ( C ) MCF10A-derived models representing non-transformed mammary stem cells (MCF10A-CT), early (MC26) and more transformed (M1B26) cells as previously described . Nuclei are stained in blue, actin filaments in red, PS in green. ( D ) Confocal images of PS and PET nanoplastic internalization in MCF10A after 24 h compared to non-exposed controls. Nuclei are stained blue actin filaments red, PS green, PET orange. ( E ) Schematic diagram of the nanoplastic (NPLs) long-term exposure approach and analysis of their impact. ( F ) Quantification of the sphere-forming ability of untreated (CT) MCF10A cells or following 20 weeks of exposure to PS or PET nanoplastics (NPLs), n=5. ( G ) Quantification of total stem cell-derived progenitors by E-CFC assay , data represent the total number of colonies per 250 seeded cells, n=5. ( F , G , ) Each individual datum is represented. One way ANOVA is indicated on the graph by the P values. ( H ) Representation of the mean proportion of myoepithelial, mixed and luminal colonies quantified by E-CFC assay (n = 5). Data are expressed as the percentage of each subtype with a total number of scored colonies representing 100%. ( I ) Confocal images of acini. Nuclei are stained in blue, actin filaments in red, white arrows point to acini with a disorganized structure. Measurement of acini ( J ) solidity or ( K ) Ferret diameter ratio was quantified by image analysis using Fiji imaging software, n=109 to 131 individual acini. ( J , K ) Each individual data is represented. Mann-Whitney test is indicated on the graph by the P values.
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Representative confocal images of fluorescent PS nanoplastic internalization after the indicated time in: ( A ) primary bone marrow CD34+ sorted hematopoietic immature cells or ( B ) <t>epithelial</t> or fibroblast cells isolated from primary mammary gland obtained from healthy donors or ( C ) MCF10A-derived models representing non-transformed mammary stem cells (MCF10A-CT), early (MC26) and more transformed (M1B26) cells as previously described . Nuclei are stained in blue, actin filaments in red, PS in green. ( D ) Confocal images of PS and PET nanoplastic internalization in MCF10A after 24 h compared to non-exposed controls. Nuclei are stained blue actin filaments red, PS green, PET orange. ( E ) Schematic diagram of the nanoplastic (NPLs) long-term exposure approach and analysis of their impact. ( F ) Quantification of the sphere-forming ability of untreated (CT) MCF10A cells or following 20 weeks of exposure to PS or PET nanoplastics (NPLs), n=5. ( G ) Quantification of total stem cell-derived progenitors by E-CFC assay , data represent the total number of colonies per 250 seeded cells, n=5. ( F , G , ) Each individual datum is represented. One way ANOVA is indicated on the graph by the P values. ( H ) Representation of the mean proportion of myoepithelial, mixed and luminal colonies quantified by E-CFC assay (n = 5). Data are expressed as the percentage of each subtype with a total number of scored colonies representing 100%. ( I ) Confocal images of acini. Nuclei are stained in blue, actin filaments in red, white arrows point to acini with a disorganized structure. Measurement of acini ( J ) solidity or ( K ) Ferret diameter ratio was quantified by image analysis using Fiji imaging software, n=109 to 131 individual acini. ( J , K ) Each individual data is represented. Mann-Whitney test is indicated on the graph by the P values.
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Representative confocal images of fluorescent PS nanoplastic internalization after the indicated time in: ( A ) primary bone marrow CD34+ sorted hematopoietic immature cells or ( B ) epithelial or fibroblast cells isolated from primary mammary gland obtained from healthy donors or ( C ) MCF10A-derived models representing non-transformed mammary stem cells (MCF10A-CT), early (MC26) and more transformed (M1B26) cells as previously described . Nuclei are stained in blue, actin filaments in red, PS in green. ( D ) Confocal images of PS and PET nanoplastic internalization in MCF10A after 24 h compared to non-exposed controls. Nuclei are stained blue actin filaments red, PS green, PET orange. ( E ) Schematic diagram of the nanoplastic (NPLs) long-term exposure approach and analysis of their impact. ( F ) Quantification of the sphere-forming ability of untreated (CT) MCF10A cells or following 20 weeks of exposure to PS or PET nanoplastics (NPLs), n=5. ( G ) Quantification of total stem cell-derived progenitors by E-CFC assay , data represent the total number of colonies per 250 seeded cells, n=5. ( F , G , ) Each individual datum is represented. One way ANOVA is indicated on the graph by the P values. ( H ) Representation of the mean proportion of myoepithelial, mixed and luminal colonies quantified by E-CFC assay (n = 5). Data are expressed as the percentage of each subtype with a total number of scored colonies representing 100%. ( I ) Confocal images of acini. Nuclei are stained in blue, actin filaments in red, white arrows point to acini with a disorganized structure. Measurement of acini ( J ) solidity or ( K ) Ferret diameter ratio was quantified by image analysis using Fiji imaging software, n=109 to 131 individual acini. ( J , K ) Each individual data is represented. Mann-Whitney test is indicated on the graph by the P values.

Journal: bioRxiv

Article Title: Chronic Exposure to Nanoplastics Alters Stem Cell Type-Specific Mechanisms, Promoting Cancer Development

doi: 10.64898/2026.01.21.700811

Figure Lengend Snippet: Representative confocal images of fluorescent PS nanoplastic internalization after the indicated time in: ( A ) primary bone marrow CD34+ sorted hematopoietic immature cells or ( B ) epithelial or fibroblast cells isolated from primary mammary gland obtained from healthy donors or ( C ) MCF10A-derived models representing non-transformed mammary stem cells (MCF10A-CT), early (MC26) and more transformed (M1B26) cells as previously described . Nuclei are stained in blue, actin filaments in red, PS in green. ( D ) Confocal images of PS and PET nanoplastic internalization in MCF10A after 24 h compared to non-exposed controls. Nuclei are stained blue actin filaments red, PS green, PET orange. ( E ) Schematic diagram of the nanoplastic (NPLs) long-term exposure approach and analysis of their impact. ( F ) Quantification of the sphere-forming ability of untreated (CT) MCF10A cells or following 20 weeks of exposure to PS or PET nanoplastics (NPLs), n=5. ( G ) Quantification of total stem cell-derived progenitors by E-CFC assay , data represent the total number of colonies per 250 seeded cells, n=5. ( F , G , ) Each individual datum is represented. One way ANOVA is indicated on the graph by the P values. ( H ) Representation of the mean proportion of myoepithelial, mixed and luminal colonies quantified by E-CFC assay (n = 5). Data are expressed as the percentage of each subtype with a total number of scored colonies representing 100%. ( I ) Confocal images of acini. Nuclei are stained in blue, actin filaments in red, white arrows point to acini with a disorganized structure. Measurement of acini ( J ) solidity or ( K ) Ferret diameter ratio was quantified by image analysis using Fiji imaging software, n=109 to 131 individual acini. ( J , K ) Each individual data is represented. Mann-Whitney test is indicated on the graph by the P values.

Article Snippet: Cells were seeded at a density of 1500 cells/mL on 96-well ultra-low-attachment plates (Corning) in serum free basal mammary epithelial medium (Promocell) supplemented with B27, 20 ng/mL epithelial growth factor (EGF), 20 ng/mL basic fibroblast growth factor (bFGF), and 4 μg/mL heparin.

Techniques: Isolation, Derivative Assay, Transformation Assay, Staining, Imaging, Software, MANN-WHITNEY