bewo cells  (ATCC)

Journal: bioRxiv

Article Title: Real-time, label-free assessment of cell fusion dynamics by high-content imaging

doi: 10.64898/2026.04.08.717136

Figure Lengend Snippet: (A-F) Representative images of BeWo cells stained fbr F-actin (Red) and nuclei (Blue). Control cells (A, C, E) exhibit discrete cell boundaries and cortical actin organization, whereas fbrskolin-treated cells (B, D, F) display loss of distinct borders and formation of multinucleated structures. Scale bars: A, B = 200 μm; C-F = 50 μm. Boxed area in A and B is magnified. (G) Quantification of F-actin fluorescence intensity in control and fbrskolin-treated cells. Data represent mean ± SEM (n = IO images per condition). Statistical significance was assessed using an unpaired two-tailed t-test (***p < 0.001).(H, 1) Relative mRNA expression of CDH1 (H) and ERVW-1 (Syncytin-1) (1) following forskolin treatment. Gene expression was normalized to 18S rRNA and expressed relative to untreated controls. Data represent mean ± SEM from three independent experiments. Statistical significance was assessed using an unpaired two-tailed t-test (*p < 0.05).

Article Snippet: BeWo human choriocarcinoma cells (ATCC) and HeLa cells (ATCC) were cultured in DMEM/F12 (1:1) medium (HiMedia) supplemented with 15% fetal bovine serum and 1% antibiotic– antimycotic solution (Gibco).

Techniques: Staining, Control, Fluorescence, Two Tailed Test, Expressing, Gene Expression

Journal: bioRxiv

Article Title: Real-time, label-free assessment of cell fusion dynamics by high-content imaging

doi: 10.64898/2026.04.08.717136

Figure Lengend Snippet: Representative bright-field time-lapse images of BeWo cells cultured under control (A) or fbrskolin-treated (B) conditions over 48h. (C, D) Automated segmentation and cluster identification using Hannony 5.1 software, with system-assigned pseudocolors indicating distinct detected clusters at the indicated time points. (E, F) Representative digital phase contrast (DPC) images illustrating intracellular texture patterns in control (E) and fbrskolin-treated (F) cells. (G-J) Quantitative time-course analysis of fusion-associated parameters measured at 6 h intervals over 48h: (G) total cluster area (×10 5 μm 2 ). (H) number of detected cell clusters, (I) area-to-cluster ratio (×10 3 ), and (J) cytoplasmic granularity quantified using the SER Bright feature. Control cells are shown in blue and fbrskolin-treated cells in red. Data represent mean ± SEM pooled from three independent experiments (n = 10 images per condition per experiment). Imaging was performed using an Operetta CLS high-content screening system.

Article Snippet: BeWo human choriocarcinoma cells (ATCC) and HeLa cells (ATCC) were cultured in DMEM/F12 (1:1) medium (HiMedia) supplemented with 15% fetal bovine serum and 1% antibiotic– antimycotic solution (Gibco).

Techniques: Cell Culture, Control, Software, Imaging, High Content Screening

Journal: bioRxiv

Article Title: Real-time, label-free assessment of cell fusion dynamics by high-content imaging

doi: 10.64898/2026.04.08.717136

Figure Lengend Snippet: BeWo cells were treated with forskolin (50 μM), a cell-permeable 8-Br-cAMP (cAMP, 1.5 μM), Wortmannin, or their respective combinations, and analyzed using the described imaging and analysis pipeline. (A-C) Quantification of area-to-cluster ratio (A), number of detected clusters (B), and cytoplasmic granularity (C) following treatment with forskolin or cAMP analog relative to untreated controls. (D-F) Corresponding parameters measured following treatment with forskolin alone, Wortmannin alone, or their combination. Data represent mean ± SEM from live-cell imaging experiments (n = 10 images per condition across two independent experiments). Imaging was performed using the Operetta CLS system and analysis was done using Hannony 5.1 software.

Article Snippet: BeWo human choriocarcinoma cells (ATCC) and HeLa cells (ATCC) were cultured in DMEM/F12 (1:1) medium (HiMedia) supplemented with 15% fetal bovine serum and 1% antibiotic– antimycotic solution (Gibco).

Techniques: Imaging, Live Cell Imaging, Software

Journal: bioRxiv

Article Title: The Epigenetic Factor PHF13 Governs Trophoblast Stemness and Differentiation

doi: 10.64898/2026.03.14.711150

Figure Lengend Snippet: A. PHF13 ko in BeWo cells, validated by western blot. Histone H3 was used as a loading control. B. PHF13 ko increases medium hCG release (ELISA) and expression (qPCR). BeWo cells were exposed to 50 mM forskolin for 72 h to induce differentiation. C. The effects of PHF13 ko on stemness genes including ELF5 , TEAD4 , and TP63 , and fusion-promoting genes, including GCM1 , ERVFRD-1 and its receptor MFSD2a . N=3, *p < 0.01, t-test.

Article Snippet: Human choriocarcinoma BeWo cell lines (ATCC, #CCL-98) were cultured in F-12K Kaighn’s modified medium (ATCC, #30-2004), supplemented with 10% FBS (ATCC, #30-2020) and 1% penicillin–streptomycin and were fed with the new F-12K complete medium every 3 days and passaged using TrypLE express enzyme, phenol red (Thermo Fisher, #12605010).

Techniques: Western Blot, Control, Enzyme-linked Immunosorbent Assay, Expressing

Journal: bioRxiv

Article Title: The Epigenetic Factor PHF13 Governs Trophoblast Stemness and Differentiation

doi: 10.64898/2026.03.14.711150

Figure Lengend Snippet: A. Gene set enrichment analysis of canonical trophoblastic progenitor genes (shown as STEMNESS, normalized enrichment score: 1.44; FDR-adjusted value: 0.07) and syncytiotrophoblast-dominant genes (shown as STB, normalized enrichment score: 2.11; FDR-adjusted value: 1 x 10 -5 ) in TS cells deficient for PHF13 . Key gene names and results are detailed in the text. B. A volcano plot showing the 621 differentially expressed mRNAs shared in both PHF13 kd lines, when compared to controls. The x-axis represents the log 2 fold change in expression (kd/ctrl), whereas the y-axis shows the minus log 10 of the FDR-adjusted p value. The horizontal dash line marks the signficance threshold (FDR-adjusted p <0.05; –log 10 > 1.30). Red points highlight downregulated genes, including PHF13, WNT6, and SOX15 , and upregulated genes that are largely expressed in differentiated trophoblasts, including ERVFRD-1, CYP19A1, LGALS16, GCM1, and CGB8. C. A volcano plot showing the 146 differentially expressed genes shared between PHF13 kd TS cells and ko BeWo cells. The x-axis represents log 2 fold change in expression (kokd/ctrl), whereas the y-axis shows the minus log 10 of the FDR-adjusted p value. The horizontal dash line marks the signficance threshold (FDR-adjusted p <0.05; –log 10 > 1.30). Red points indicate significantly altered genes, which include downregulated stemness-associated genes ( WNT6 and SOX15 ), and upregulated differentiated trophoblast markers (e.g. ERVFRD-1, CYP19A1, LGALS16, GCM1, CGA, CGB5, CGB3, CGB8, and PGF ).

Article Snippet: Human choriocarcinoma BeWo cell lines (ATCC, #CCL-98) were cultured in F-12K Kaighn’s modified medium (ATCC, #30-2004), supplemented with 10% FBS (ATCC, #30-2020) and 1% penicillin–streptomycin and were fed with the new F-12K complete medium every 3 days and passaged using TrypLE express enzyme, phenol red (Thermo Fisher, #12605010).

Techniques: Expressing

Journal: bioRxiv

Article Title: The Epigenetic Factor PHF13 Governs Trophoblast Stemness and Differentiation

doi: 10.64898/2026.03.14.711150

Figure Lengend Snippet: A-B. 37 downregulated genes and 48 upregulated genes in PHF13-deficient cells are direct targets of PHF13. Note that their enrichment is signficant with 2.11-fold and 2.93-fold enrichment, respectively. C. PHF13 binds genomic regions of IFITM3 , an inhibitor of trophoblast fusion, in TS-CTB but not in TS-STB. Yellow shading indicates significant PHF13 peaks relative to IgG control. The x-axis represents genomic corordinates with exonic and intronic structure, and arrows indicate transcription direction. The y-axis represents normalized peak intensity. D-E. PHF13 promotes the expression of IFITM3 transcripts (D) and protein (E) in PHF13 ko BeWo cells. Actin was used as a loading control. N=3, * p < 0.05, t-test.

Article Snippet: Human choriocarcinoma BeWo cell lines (ATCC, #CCL-98) were cultured in F-12K Kaighn’s modified medium (ATCC, #30-2004), supplemented with 10% FBS (ATCC, #30-2020) and 1% penicillin–streptomycin and were fed with the new F-12K complete medium every 3 days and passaged using TrypLE express enzyme, phenol red (Thermo Fisher, #12605010).

Techniques: Control, Expressing