Journal: Cell Biology International

Article Title: Tomatidine Exposure During Early Myogenesis Promotes Hypertrophy of C2C12 Muscle Cells Through Increased Nuclei Accretion

doi: 10.1002/cbin.70224

Figure Lengend Snippet: Exposure of muscle cells to tomatidine during proliferation reduces C2C12 myoblast proliferation. C2C12 cells were grown for 3 days in standard growth media containing vehicle control (DMSO) or media supplemented with 10 µM tomatidine, in the xCELLigence Instrument. (A) Schematic of experimental timeline. Created in BioRender. Caldow, M. (2026) https://BioRender.com/paic1qg . (B) Growth curve of Cell Index over 72 h of treatment time, calculated via electric impedance measurements on the xCELLigence Instrument. (C) Area under the curve of Cell Index data. (D) Mean doubling time (Td), calculated from electric impedance measurements on the xCELLigence. Data are presented as mean ± SD ( n = 3 experiments). Significantly different to DMSO, ** p < 0.01, *** p < 0.001. DMSO, vehicle supplemented control cells. TOM, cells supplemented with 10 µM tomatidine.

Article Snippet: Murine C2C12 myoblasts (ATCC Cryosite distribution, NSW, Australia) were cultured in growth media [GM, DMEM (#11965; ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) antibiotic solution (100 unit/ml penicillin/streptomycin) (Life Technologies, ThermoFisher Scientific)] at 37°C in an atmosphere of 5% CO 2 .

Techniques: Control

Journal: Cell Biology International

Article Title: Tomatidine Exposure During Early Myogenesis Promotes Hypertrophy of C2C12 Muscle Cells Through Increased Nuclei Accretion

doi: 10.1002/cbin.70224

Figure Lengend Snippet: Muscle cells supplemented with tomatidine during early differentiation exhibit hypertrophy and increased cell fusion. C2C12 cells were differentiated for 2 days in standard differentiation media containing vehicle control (DMSO) or media supplemented with 10 µM tomatidine from the onset of differentiation. (A) Schematic of experimental timeline. Created in BioRender. Caldow, M. (2026) https://BioRender.com/8nz1y75 . (B) Representative images of C2C12 cells differentiated for 2 days in standard differentiation media containing vehicle control (DMSO) or media supplemented with 10 µM tomatidine from the onset of differentiation. Cells were immunostained for myosin (red) and DAPI (blue, nuclei). (C) Myotube diameter. (D) Number of multinucleated MF20+ cells. (E) Number of nuclei. (F) Number of nuclei within multinucleated MF20+ cells. (G) Fusion index. (H) Differentiation index. (I) Puromycin protein abundance and representative immunoblot. Data are presented as mean ± SD ( n = 3 experiments), normalized to DMSO. Significantly different to DMSO, ** p < 0.01, *** p < 0.001. GM, growth media before differentiation media was applied. DMSO, vehicle treated control cells. TOM, cells supplemented with 10 µM tomatidine. Scale bar represents 50 µm.

Article Snippet: Murine C2C12 myoblasts (ATCC Cryosite distribution, NSW, Australia) were cultured in growth media [GM, DMEM (#11965; ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) antibiotic solution (100 unit/ml penicillin/streptomycin) (Life Technologies, ThermoFisher Scientific)] at 37°C in an atmosphere of 5% CO 2 .

Techniques: Control, Quantitative Proteomics, Western Blot

Journal: Cell Biology International

Article Title: Tomatidine Exposure During Early Myogenesis Promotes Hypertrophy of C2C12 Muscle Cells Through Increased Nuclei Accretion

doi: 10.1002/cbin.70224

Figure Lengend Snippet: The growth promoting effects of tomatidine supplementation on C2C12 myotubes are maintained even after returning to differentiation media. (A) C2C12 cells were differentiated for 4 days in standard differentiation media containing vehicle control (DMSO) or media supplemented with 10 µM tomatidine from the onset of differentiation. Cells were also supplemented with 10 µM tomatidine from differentiation Day 0 to Day 2, then returned to vehicle treatment media from Day 2 to Day 4 (TOM > DMSO). Or cells were supplemented with tomatidine from differentiation day 2 to day 4 only (DMSO > TOM). Created in BioRender. Caldow, M. (2026) https://BioRender.com/foud55w . (B) Representative images of cells immuno‐stained for myosin (red) and DAPI (blue, nuclei). (C) Myotube diameter. (D) Number of multinucleated MF20+ cells. (E) Number of nuclei. (F) Number of nuclei within multinucleated MF20+ cells. (G) Fusion index. (H) Differentiation index. (I) Puromycin protein abundance and representative immunoblot. Data are presented as mean ± SD ( n = 3 experiments), normalized to DMSO. Significantly different to DMSO, ** p < 0.01, *** p < 0.001. Scale bar represents 50 µm. GM, growth media before differentiation media was applied. DMSO, vehicle treated control cells. TOM, cells supplemented with 10 µM tomatidine.

Article Snippet: Murine C2C12 myoblasts (ATCC Cryosite distribution, NSW, Australia) were cultured in growth media [GM, DMEM (#11965; ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) antibiotic solution (100 unit/ml penicillin/streptomycin) (Life Technologies, ThermoFisher Scientific)] at 37°C in an atmosphere of 5% CO 2 .

Techniques: Control, Staining, Quantitative Proteomics, Western Blot

Journal: Cell Biology International

Article Title: Tomatidine Exposure During Early Myogenesis Promotes Hypertrophy of C2C12 Muscle Cells Through Increased Nuclei Accretion

doi: 10.1002/cbin.70224

Figure Lengend Snippet: Tomatidine supplementation promotes myotube‐myotube fusion. C2C12 cells were differentiated for 4 days in standard differentiation media containing vehicle control (DMSO) or media supplemented with 10 µM tomatidine from the onset of differentiation. (A) Live cell imaging was performed from D2 to D4 of differentiation. Images were captured each hour, across 45 h. Figure created with BioRender. Caldow, M. (2026) https://BioRender.com/u1n0wm7 . (B) Representative brightfield images of cells over time. (C) Schematic explaining differentiation under normal conditions (i), effects of tomatidine supplementation to promote hypertrophy via increased myonuclei‐myotube fusion (ii) or (iii) myotube‐myotube fusion. Created in BioRender. Caldow, M. (2026) https://BioRender.com/hpcc0s9 . Scale bar represents 200 µm. DMSO, vehicle treated control cells. TOM, cells supplemented with 10 µM tomatidine.

Article Snippet: Murine C2C12 myoblasts (ATCC Cryosite distribution, NSW, Australia) were cultured in growth media [GM, DMEM (#11965; ThermoFisher Scientific, Waltham, MA, USA) supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) antibiotic solution (100 unit/ml penicillin/streptomycin) (Life Technologies, ThermoFisher Scientific)] at 37°C in an atmosphere of 5% CO 2 .

Techniques: Control, Live Cell Imaging, Brightfield

Journal: Advanced Science

Article Title: Ultrafast Micrometric Laser Structuring of Polymer Surfaces for Highly Organized Skeletal Muscle Tissue Modeling

doi: 10.1002/advs.78129

Figure Lengend Snippet: Relative gene expression of C2C12 myoblasts cultured on the LS surface compared with the standard surface. Data are presented as mean ± SD with individual data points ( n = 3). Gene expression levels were normalized to the housekeeping gene and expressed as fold change relative to the standard group (Std). Panels correspond to: (a) d0, (b) d4, (c) d7, and (d) d11. Statistical significance was determined using Student's t ‐test: * p < 0.05; ** p < 0.01; *** p < 0.001 vs. Standard.

Article Snippet: Murine myoblasts C2C12 (ATCC) were cultured according to a previously published protocol [ ] in T‐75 Flasks (Sarstedt) at 37°C and 5% CO 2 in Dulbecco's modified Eagle medium GlutaMAX (DMEM, Gibco) supplemented with 10% heat‐inactivated fetal bovine serum (FBS, EuroClone), penicillin (100 IU mL −1 , Gibco), and streptomycin (100 mg mL −1 , Gibco).

Techniques: Gene Expression, Cell Culture

Journal: Advanced Science

Article Title: Ultrafast Micrometric Laser Structuring of Polymer Surfaces for Highly Organized Skeletal Muscle Tissue Modeling

doi: 10.1002/advs.78129

Figure Lengend Snippet: (a) Infrared spectra of the standard surface before (Std, red line) and after laser structuring (LS, blue line); (b) MTT assay. The C2C12 cell growth at three experimental time points (24 h, 48 h, and 72 h) is plotted for the cells seeded on a standard surface (Std, red line) and LS (ex. LS, blue line) surfaces. A theoretical curve was also plotted, taking into account the area available on the LS surface (th. LS, dashed dark blue line). (a.u. = arbitrary unit); (c) Brightfield pictures of the cells cultured on standard (top pictures) or LS (bottom pictures) surfaces at the three experimental time points (24 h, 48 h, and 72 h).

Article Snippet: Murine myoblasts C2C12 (ATCC) were cultured according to a previously published protocol [ ] in T‐75 Flasks (Sarstedt) at 37°C and 5% CO 2 in Dulbecco's modified Eagle medium GlutaMAX (DMEM, Gibco) supplemented with 10% heat‐inactivated fetal bovine serum (FBS, EuroClone), penicillin (100 IU mL −1 , Gibco), and streptomycin (100 mg mL −1 , Gibco).

Techniques: Infrared, MTT Assay, Brightfield, Cell Culture

Journal: Advanced Science

Article Title: Ultrafast Micrometric Laser Structuring of Polymer Surfaces for Highly Organized Skeletal Muscle Tissue Modeling

doi: 10.1002/advs.78129

Figure Lengend Snippet: Image analyses conducted on C2C12 after 7 days of differentiation. Pictures a, b, and c show representative immunofluorescence pictures superimposed on the top with their relative brightfield pictures. The myotubes positive for MHC (Red), are randomly oriented in the standard plates (a) while properly aligned on the LS surfaces (channel spacing of 75 µm (b) and 150 µm (c)). Orientation of muscle fibers. The distribution of orientation is plotted for unstructured standard plates (Std) and for all spacing (d, 75, 100, 125, and 150 µm) and depth (g, 3.5 – 5 – 8;5 – 11.5 – 14 – 15 ‐20 µm) values tested, revealing the presence of a prevalent peak for all conditions compared to their respective control (black plot); the quantification of the orientation through the coherency value (e, f) shows a significative increase for all the LS surfaces compared to the standard ( * ). Moreover, regarding the spacing analysis, the three higher values are also significative compared to 75 µm (e, $ ). Myotube lengths. The myotubes differentiated on LS surfaces with a spacing of 125 µm and a depth of 11.5 µm (h, red bar) are significantly longer than those grown on the standard plates (h, black bar). Sample size n = 3. p value list: spacing, **** p < 0.0001, *** p = 0.0001, $$$ p = 0.0006, $$ p = 0.0013 (75 µm vs 100 µm), $$ p = 0.0059 (75 µm vs 150 µm); depth, **** p < 0.0001, *** p = 0.0007, ** p = 0.001 (3.5 µm), ** p = 0.0025 (14 µm), ** p = 0.0085 (20 µm); length, **** p < 0.0001.

Article Snippet: Murine myoblasts C2C12 (ATCC) were cultured according to a previously published protocol [ ] in T‐75 Flasks (Sarstedt) at 37°C and 5% CO 2 in Dulbecco's modified Eagle medium GlutaMAX (DMEM, Gibco) supplemented with 10% heat‐inactivated fetal bovine serum (FBS, EuroClone), penicillin (100 IU mL −1 , Gibco), and streptomycin (100 mg mL −1 , Gibco).

Techniques: Immunofluorescence, Brightfield, Control, Analysis

Journal: Advanced Science

Article Title: Ultrafast Micrometric Laser Structuring of Polymer Surfaces for Highly Organized Skeletal Muscle Tissue Modeling

doi: 10.1002/advs.78129

Figure Lengend Snippet: Analysis of MHC and Desmin protein levels during C2C12 myoblast differentiation. Western blot analysis of C2C12 cells cultured on standard (Std) and LS surfaces at different stages of differentiation. Representative immunoblot for (a) MHC and Tubulin, and (c) Desmin and GAPDH. Densitometric analysis of (b) MHC and (d) Desmin protein levels. Data are presented as mean ± SD with individual data points ( n = 3). Statistical significance was assessed using Student's t ‐test ( * p < 0.05; ** p < 0.01; *** p < 0.001 vs. Standard at each time point).

Article Snippet: Murine myoblasts C2C12 (ATCC) were cultured according to a previously published protocol [ ] in T‐75 Flasks (Sarstedt) at 37°C and 5% CO 2 in Dulbecco's modified Eagle medium GlutaMAX (DMEM, Gibco) supplemented with 10% heat‐inactivated fetal bovine serum (FBS, EuroClone), penicillin (100 IU mL −1 , Gibco), and streptomycin (100 mg mL −1 , Gibco).

Techniques: Analysis, Western Blot, Cell Culture