Journal: Frontiers in Cell and Developmental Biology

Article Title: Optimised dissociation and multimodal profiling of prostate cancer stroma reveal fibromuscular cell heterogeneity with clinical correlates

doi: 10.3389/fcell.2025.1653780

Figure Lengend Snippet: Viable cell yields differ widely between distinct tissue dissociation protocols. (A) Schematic comparison of the tissue dissociation protocols employed demonstrating: common tissue pre-processing steps; enzymatic dissociation with enzyme cocktail composition and incubation times; post-enzymatic dissociation downstream processing steps; and quality assessment via cell viability analysis per trypan blue staining and cell seeding. Readouts are exemplified in (B–D) . (B–E) Analysis of benign tissue cores dissociated using the indicated protocol. (B) Viable cell yield/mg tissue for each protocol. Bars represent the median ± interquartile range from 5, 6 or 29 biological replicates for the Miltenyi and BD 30 min, BD 60 min or optimised protocols, respectively. Statistical significance was determined using Brown-Forsythe and Welch ANOVA tests with Dunnett T3 correction for multiple comparisons. (C) Brightfield imaging for trypan blue viability assessment of freshly dissociated cells isolated using the indicated protocol. Scale bars represent 200 μm at ×10 magnification. (D) Brightfield images of the 10% seeded cells after 4 weeks. Scale bars represent 500 μm at ×4 magnification. Frequency of successful mesenchymal cell growth for each of the four replicates is indicated. (E) Immunofluorescent staining of successfully cultured cells for the epithelial marker pan-cytokeratin (panCK) and mesenchymal markers vimentin and CD90, whereby font colour denotes pseudocolouring in the displayed images. Nuclei were counterstained using Hoechst 33342 (blue). 22Rv1 prostate cancer cells and explant cultures of primary human prostate fibroblasts served as negative and positive controls for panCK or vimentin and CD90, respectively. Scale bars represent 100 μm at ×20 magnification.

Article Snippet: The 22Rv1 prostate cancer cell line was obtained from the American Type Culture Collection (ATCC Nr. CRL-2505; ATCC; Rockville, MD), STR validated and maintained according to the distributor’s instructions.

Techniques: Comparison, Incubation, Staining, Imaging, Isolation, Cell Culture, Marker

Journal: Oncogene

Article Title: Effect of TGF-β mediated phenotypic changes on prostate cancer cell anoikis response

doi: 10.1038/s41388-025-03600-z

Figure Lengend Snippet: a Prostate cancer cell lines were subjected to treatment with CBZ (10 nM) alone or in combination with DZ-50 (2 or 5 μM) for 24, 48 or 72 h. C42B-TaxR cells that are unresponsive to CBZ die in response to DZ-50. An improved therapeutic response is observed with combination therapy of DZ-50 and CBZ (C + D) in 22RV1, LNCaP, LNCaPTRII, and C42BTaxR cells. Bars ± SEM. * P < 0.05, ** p < 0.01 *** p < 0.001, **** p < 0.0001. n = 3 independent experiments. Each experiment was performed in triplicates. b Cell viability assessment in CRPC organoids after 72 h treatment with six doses as indicated of DZ-50 alone or CBZ (10 nM) in combination with DZ-50.

Article Snippet: Human prostate cancer cell lines 22RV1 (CRPC) (CRL-2505) (RRID:CVCL_1045) and LNCaP (androgen sensitive) (CRL-1740) (RRID:CVCL_1379) were obtained from ATCC, Manassas, VA. Therapeutically resistant human prostate cancer cell lines (C4-2B parental and C4-2B TaxR -TGFβ unresponsive, taxane resistant) were generously provided by Dr. Allen Gao (University of California, Davis) [ , ] (C4-2B parental: RRID: CVCL_4784, C4-2B TaxR: developed in Gao lab by culturing C4-2B cells in docetaxel in a dose-escalation manner).

Techniques: Clinical Proteomics