Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Standalone methacrylated extracellular matrix for digital light processing bioprinting: a practical workflow

doi: 10.3389/fbioe.2026.1774476

Figure Lengend Snippet: 3D bioprinted dECM-MA bioink supports high-density parenchymal cell attachment, and early ECM remodeling. Human uterine stromal fibroblast cells (primary parenchymal cells of the uterine tissue, FC-0076 Lifeline Cell Technologies) were cultured in commercial media, exhibiting characteristic spindle-shaped stromal morphology (A) . These cells were immunofluorescently characterized for positive uterine and stromal markers (CD10, CD73, and Vimentin, respectively). CD31 isotype staining served as a negative control. Following expansion, cells were seeded at high density onto 3D bioprinted dECM-MA scaffolds, cultured for 16 h, and characterized by HR-SEM (B) . An overview SEM image shows a visibly high cell density on the ECM-MA after culture. Higher magnifications reveal a high-density (yellow rectangle, zoomed in) and lower density (blue rectangle, zoomed in) areas where human uterine stromal cells (yellow arrow) interact with the dECM-MA fibers (green arrow), suggesting biological interaction and constructive remodeling.

Article Snippet: Specifically, rabbit anti-human CD10 (Bioss, Cat. No. BS-0527R-20; RID: AB_10854297) and rabbit anti-human vimentin (Bioss, Cat. No. BS-0756R-20; RRID: AB_10855343) were used as primary antibodies, with a rabbit IgG isotype control (Santa Cruz Biotechnology, Cat. No. sc-8306; RRID: AB_653100).

Techniques: Cell Attachment Assay, Cell Culture, Staining, Negative Control

Journal: Journal of Translational Medicine

Article Title: NGF-mediated tumor–stroma crosstalk promotes prostate cancer aggressiveness

doi: 10.1186/s12967-026-07915-7

Figure Lengend Snippet: PC-derived CAFs release NGF and express mesenchymal/myofibroblast markers as well as TrkA. ( A ) Quantification of NGF secretion by ELISA in conditioned media collected from primary CAFs lines (C1–C8) isolated from independent PC patients at Gleason’s scores 6–8. Data are expressed as pg/ml (mean ± SD) from three independent experiments performed in duplicate. ( B ) Representative IF images of NGF expression in primary CAFs (C1). NGF is shown in green and nuclei in blue. Images are representative of three independent experiments. Scale bar, 10 µM. ( C ) Quantitative analysis of NGF fluorescence intensity in primary CAF cultures (C1–C8). Mean fluorescence intensity (MFI) was normalized to the number of nuclei and expressed as fold increase relative to the fluorescence from the secondary antibody, used as control (II Ab). Data are presented as mean ± SD from several independent fields analyzed for each culture. ( D ) Western blot analysis of TrkA and mesenchymal/myofibroblast markers in lysates from primary prostate CAFs (C1–C8). Vimentin was used as a mesenchymal marker, while α-SMA and FAP-1 were used as myofibroblast-associated markers. Tubulin served as loading control. LNCaP cells, positive for the expression of E-cadherin, were included as epithelial control

Article Snippet: The following primary antibodies were used: mouse monoclonal antibodies against α-tubulin (#E-AB-20036, Elabsciences, Houston, TX, USA), GAPDH (#E-AB-20079, Elabsciences), α-SMA (CGA7, Santa Cruz Biotechnology, Dallas, TX, USA), FAP (SS-13, Santa Cruz Biotechnology), and E-cadherin (Clone 36, RUO); rabbit polyclonal antibodies against vimentin (H-84, Santa Cruz Biotechnology, Dallas, TX, USA) and TrkA (06-574, Millipore, Burlington, MA, USA).

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Isolation, Expressing, Fluorescence, Control, Western Blot, Marker

Journal: Journal of Translational Medicine

Article Title: NGF-mediated tumor–stroma crosstalk promotes prostate cancer aggressiveness

doi: 10.1186/s12967-026-07915-7

Figure Lengend Snippet: Soluble factors derived from CAFs promote spheroid growth and induce EMT-like changes in PC-derived spheroids cultured in 3D. ( A ) Representative phase-contrast and immunofluorescence (IF) images showing vimentin expression (red) in LNCaP-derived spheroids cultured for 15 days in control medium or CAF-derived conditioned medium (CAF-CM). ( B ) Quantification of LNCaP spheroid size expressed as fold increase relative to control conditions after 15 days of treatment with ctrl medium or CAF-derived CM ( n = 3). Bar, 100 µM. Data are presented as mean ± SD. ** p < 0.01. ( C ) Western blot analysis of EMT markers (E-cadherin, vimentin) in LNCaP cells cultured in 2D or 3D conditions in the absence or presence of CAF-CM and anti-NGF neutralizing Ab (anti-NGF Ab). ( D ) Densitometric quantification of vimentin/tubulin and ( E ) E-cadherin/tubulin ratios from independent experiments shown in ( C ), including conditions with anti-NGF neutralizing antibody (anti-NGF Ab) where indicated. ( F ) Representative phase-contrast and immunofluorescence images showing vimentin expression (red) in PC3-derived spheroids cultured for 15 days in control medium or CAF-CM. ( G ) Quantification of PC3 spheroid size, expressed as fold increase relative to control conditions after 15 days of treatment ( n = 3). Data are presented as mean ± SD. ** p < 0.01. Bar 100 µM. ( H ) Western blot analysis of EMT markers (E-cadherin and vimentin) in PC3 cells cultured under 2D or 3D conditions. ( I-L ) Densitometric analysis of vimentin/tubulin and E-cadherin/tubulin ratios from experiments shown in ( H )

Article Snippet: The following primary antibodies were used: mouse monoclonal antibodies against α-tubulin (#E-AB-20036, Elabsciences, Houston, TX, USA), GAPDH (#E-AB-20079, Elabsciences), α-SMA (CGA7, Santa Cruz Biotechnology, Dallas, TX, USA), FAP (SS-13, Santa Cruz Biotechnology), and E-cadherin (Clone 36, RUO); rabbit polyclonal antibodies against vimentin (H-84, Santa Cruz Biotechnology, Dallas, TX, USA) and TrkA (06-574, Millipore, Burlington, MA, USA).

Techniques: Derivative Assay, Cell Culture, Immunofluorescence, Expressing, Control, Western Blot