Journal: Scientific Reports

Article Title: Changes in ovarian hardness and elasticity affect the development and function of secondary follicles

doi: 10.1038/s41598-026-39396-5

Figure Lengend Snippet: Effects of oocyte-secreted factors on secondary follicles cultured in 0.1 % alginate bead with viscosity 4-12 cP. A; The effect of oocyte-secreted factors on the development of secondary follicles with multilayered granulosa cells. Ten secondary follicles with multilayered granulosa cells were placed in each alginate bead and then were cultured for 7 days in culture medium containing 10 ng/ml ovine FSH, GDF9 (0, 10 and 100 ng/ml) and ITS Supplement in the presence of 1 % (v/v) FBS. The diameter of each secondary follicle was measured 7 days after culture. In the box plot: bar, × = median, box = 25 th to 75 th percentiles, whiskers = 10 th and 90 th percentile. N=3 animals for each treatment. The experiment was repeated three times. The different symbols represent significant differences (p < 0.05). *; Significant differences observed between days 0 and 7 in the 10 and 100 ng/mL GDF9 groups (p < 0.05). B; The effect of oocyte-secreted factors on the expression of marker genes about the follicular development, luteinization and inflammation/anti-inflammatory in granulosa cells of cultured secondary follicles. Ten secondary follicles with multilayered granulosa cells were placed in each alginate bead and then were cultured for 7 days in culture medium containing 10 ng/ml ovine FSH, GDF9 (0, 10 and 100 ng/ml) and ITS Supplement in the presence of 1 % (v/v) FBS. The diameter of each secondary follicle was measured 7 days after culture. Levels of mRNA were normalized to that of L19 . The values are the mean ± SEM. N=3 animals for each treatment. The experiment was repeated three times. The value of 0 ng/ml GDF9 was set as 1, and the data are presented as fold induction. *Significant differences observed between the 0 ng/mL and 10 or 100 ng/mL GDF9 groups (p < 0.05).

Article Snippet: Ten of secondary follicles were placed in one alginate bead and were cultured for 7 days in the medium (DMEM/F12 containing penicillin and streptomycin) containing 10 ng/ml ovine FSH (NIDDK, Torrance, CA, USA) and ITS Supplement (Sigma) in the presence of 1 % (v/v) FBS with or without mouse recombinant GDF9 (0, 10 and 100 ng/ml) (R&D Systems Inc., Minneapolis, MN) or Cytochalasin (0, 1 and 10 μM) (Sigma) at 37 °C and 5% CO 2 condition.

Techniques: Cell Culture, Viscosity, Expressing, Marker

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: Dysregulation of BMP9/ALK1 signaling and inflammation in refractory ulcerative colitis (rUC). ( A ) Heatmap depicting serum expression levels of BMP family members in healthy controls, non-rUC, and rUC patients ( n = 3 per group). Data were Z-score normalized and hierarchically clustered (red, high expression; blue, low expression). ( B ) Baseline serum levels of BMP9 and BMP10 in healthy controls ( n = 50), non-rUC patients ( n = 47), and rUC patients ( n = 48). ( C ) Spearman correlation analyses between baseline serum BMP9 levels and clinical disease activity indices, including baseline Modified Mayo Score, baseline UCEIS, post-treatment Modified Mayo Score, and post-treatment UCEIS, in patients with UC ( n = 95). ( D ) Colonic mucosal mRNA expression levels of ALK1, IL-6, TNF-α, and CCL2 in healthy controls, non-rUC, and rUC patients ( n = 4 per group). Statistical annotations for ( B , D ): (Normalized to GAPDH; Mean ± SD; Statistical significance determined by one-way ANOVA with Tukey’s post hoc test: ** p < 0.01, *** p < 0.001, ns: not significant).

Article Snippet: Recombinant murine BMP9 (MCE, HY-P700530) was administered via intraperitoneal injection (200 ng/mouse/day) from day 0 to day 7, concurrent with DSS induction.

Techniques: Expressing, Activity Assay, Modification

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9 attenuates DSS-induced colitis in mice. ( A ) Schematic of experimental design: Acute colitis was induced in C57BL/6 mice by 3% DSS in drinking water for 7 days. The BMP9 treatment group received intraperitoneal injections of recombinant murine BMP9 (200 ng/day), while the DSS group and the control group received PBS ( n = 14 per group). ( B ) Serum BMP9(ng/mL) concentrations measured by ELISA. ( C ) Representative images of colons and quantitative analysis of colon length (cm). ( D ) Colonoscopy images (upper), H&E-stained colon sections (lower; scale bars = 100 μm), and histopathological scores. ( E ) (Upper) Dynamic body weight changes and (Lower) Disease Activity Index (DAI) scores. Data expressed as mean ± SD; * p < 0.05, two-way repeated measures ANOVA. ( F ) (Left) Relative Alk1 mRNA expression in colon tissues (RT-qPCR normalized to Gapdh). (Right) ALK1 protein concentrations (quantified by ELISA). Data presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 ns: not significant). ( G ) Relative mRNA expression levels of IL-1β, Ccl2, Col1a1, and Col3a1 (RT-qPCR; mean ± SD). ( H ) Western blot analysis of CCL2, TGF-β, and α-SMA protein expression in colon tissues and quantitative analysis of band intensities. ( I ) Western blot detection of p-Smad1, total Smad1, and VE-cadherin proteins and quantitative analysis of band intensities.

Article Snippet: Recombinant murine BMP9 (MCE, HY-P700530) was administered via intraperitoneal injection (200 ng/mouse/day) from day 0 to day 7, concurrent with DSS induction.

Techniques: Recombinant, Control, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9 restores intestinal vascular barrier integrity in DSS-induced colitis. ( A ) Representative immunofluorescence images of VE-cadherin (red) and CD31 (green) co-localization in colon tissues (nuclei counterstained with DAPI). ( B ) Left: Schematic of FITC-dextran (4 kDa) permeability assay. Right: Quantified serum FITC fluorescence intensity 60 min post-gavage ( n = 5 per group). ( C ) Left: Schematic of Evans Blue vascular leakage assay. Right: Colonic Evans Blue extravasation quantified by absorbance at 620 nm (mean ± SD; n = 5 per group; ** p < 0.01, *** p < 0.001, ns: not significant; one-way ANOVA with Tukey’s test). ( D ) KEGG pathway analysis of RNA-seq data from colon tissues (DSS + BMP9 groups vs. DSS). ( E ) Volcano plot of differentially expressed genes. Genes highlighted in bold are key IBD-associated downregulated factors.

Article Snippet: Recombinant murine BMP9 (MCE, HY-P700530) was administered via intraperitoneal injection (200 ng/mouse/day) from day 0 to day 7, concurrent with DSS induction.

Techniques: Immunofluorescence, Permeability, Fluorescence, RNA Sequencing

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9/ALK1 signaling modulates neutrophil migration and endothelial tube formation. ( A ) Schematic representation of the neutrophil migration assay. ( B ) (Left) Representative fluorescence micrographs demonstrating Calcein-AM-labeled neutrophil migration through 3 μm pore Transwell inserts toward HIMECs pretreated for 2 h with: vehicle control (0 ng/mL BMP9), BMP9 (0.1, 1, or 10 ng/mL), TNF-α (20 ng/mL as positive control), or ALK1 inhibitor ML347 (150 nM). (Right) Quantitative analysis of neutrophil migration rates (mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, ns: not significant by two-way ANOVA with post hoc testing). ( C ) Schematic illustration of the endothelial tube formation assay protocol. ( D ) Representative phase-contrast images of tubular network formation by HIMECs cultured on growth factor-reduced Matrigel under various treatment conditions: vehicle control (0 ng/mL BMP9), BMP9 (0.1, 1, or 10 ng/mL), TNF-α (20 ng/mL), or ML347 (150 nM). ( E ) Quantitative assessment of angiogenic parameters including branch points and nodal junctions (mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001 by two-way ANOVA with appropriate post hoc comparisons).

Article Snippet: Recombinant murine BMP9 (MCE, HY-P700530) was administered via intraperitoneal injection (200 ng/mouse/day) from day 0 to day 7, concurrent with DSS induction.

Techniques: Migration, Fluorescence, Labeling, Control, Positive Control, Endothelial Tube Formation Assay, Cell Culture

Journal: Science Advances

Article Title: Repair of female reproductive function by GDF-9–overexpressing extracellular vesicles via ACVR1B/SMAD2 regulation in ovarian granulosa

doi: 10.1126/sciadv.adw9006

Figure Lengend Snippet: ( A ) Schematic illustration of the in vitro DO model; concentration of cisplatin, 2.7 μg/ml; EV/GOE, 1 × 10 6 particles/ml; time of treatment, 24 hours (h). ( B ) Flow cytometry analysis of GC viability, apoptosis, and cell death. ( C to E ) Quantification of flow cytometry data, showing the efficacy of EVs in treating DO in vitro. ( F ) No statistically significant difference in apoptosis rates between GOEs and GDF-9 –overexpressing MSCs. n = 3, data are presented as means ± SD. This figure was created in part using BioRender.com . Created in BioRender. Zhang, S. (2025) https://BioRender.com/zv1tbzt . IC 50 , median inhibitory concentration. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.

Article Snippet: The protein concentration of GDF-9 in control EVs and GOE was quantitatively analyzed using the ELISA kit (Raybiotech, ELH-GDF9-1).

Techniques: In Vitro, Concentration Assay, Flow Cytometry

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 1. Transzonal projections (TZP) density of COCs during IVM with BMP15 (A) and/or GDF9 (B). Phalloidin-FITC fluorescence intensity (mean ± S.E.M.) in the zona pellucida area of COCs after 0h, 6h, 12h and 24h of IVM. A total of 20–30 COCs per group were assessed in 3 replicates. (A) Values with different letters differ significantly (p < 0.05). (B) Different letters between groups in the same timepoint (a,b) and different symbols between timepoints in the same group (α,β,γ for control and GDF9 groups; and σ,τ for GDF9+BMP15 group) indicate significant differences (p < 0.05). (C) Representative confocal images of phalloidin-FITC staining. TZP are observed as thin filaments going through the zona pellucida from the cumulus cells to the oocyte. Scale bar: 100 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Fluorescence, Control, Staining

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 2. Mitochondrial activity of MII-oocytes after IVM with BMP15 and/or GDF9. (A) Relative fluorescence intensity (mean ± S.E.M.) of Mitotracker stained oocytes. Columns with different letters differ significantly (p < 0.05). A total of 20–27 oocytes per group were assessed in 3 replicates. (B) Representative images. Scale bar: 100 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Activity Assay, Fluorescence, Staining

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 3. ROS and GSH levels after IVM with BMP15 and/or GDF9. Relative fluorescence intensity (mean ± S.E.M.) of (A) ROS and (B) GSH. A total of 24–31 oocytes per group were assessed in 3 replicates. (C) Representative images. Scale bar: 200 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Fluorescence

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 4. EGFR expression of MII-oocytes after IVM with BMP15 (A) and/or GDF9 (B). Relative fluorescence intensity (mean ± S.E.M.) of EGFR immunostained oocytes. Columns with different letters differ significantly (p < 0.05). A total of 26–40 oocytes per group were assessed in 4 replicates. (C) Representative images. Scale bar: 30 μm.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Expressing, Fluorescence

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 5. EGFR expression of cumulus cells after IVM with BMP15 (A) and/or GDF9 (B). Relative EGFR expression (mean ± S.E.M.). Columns with different letters differ significantly (p < 0.05). Four independent blots were used for relative quantification. (C) Western blot analyses of EGFR expression. EGFR from 70 to 80 COCs/ replicate was quantified by western blotting and standardized to vinculin protein levels.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Expressing, Quantitative Proteomics, Western Blot

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 6. Cumulus expansion index after IVM with BMP15 (A) and/or GDF9 (B). Cumulus expansion of each COC was scored on a 0 to 4 scale and cumulus expansion index (mean ± S.E.M) was calculated as the arithmetic mean value scored of all COCs evaluated per replica. Columns with different letters differ significantly (p < 0.05). A total of 367–441(A) and 278–323 (B) oocytes per group were assessed in 5 replicates.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques:

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 7. Effect of IVM supplementation with BMP15 (A) and/or GDF9 (B) on 8-day blastocyst quality after parthenogenic activation (PA). Cell number (mean ± S.E. M.) of the inner cell mass (ICM) and the trophectoderm (TE) of 8-days blastocysts. Columns with different letters differ significantly (p < 0.05). A total of 15–29 blastocysts per group were assessed.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: Activation Assay

Journal: Theriogenology

Article Title: Effect of BMP15 and GDF9 in the IVM medium on subsequent oocyte competence and embryo development of prepubertal goats.

doi: 10.1016/j.theriogenology.2024.12.015

Figure Lengend Snippet: Fig. 8. Effect of IVM supplementation with BMP15 (A) and/or GDF9 (B) on 8-day blastocyst quality after IVF. Cell number (mean ± S.E.M.) of 8-day blastocysts in the inner cell mass (ICM) and the trophectoderm (TE). Columns with different letters differ significantly (p < 0.05). A total of 14–22 blastocysts per group were assessed.

Article Snippet: The IVM medium was supplemented with 100 ng/mL of recombinant human mature region BMP15 (R&D systems, Minnesota, USA) and/or 100 ng/mL of recombinant human mature region GDF9 (R&D systems), according to the experimental design, at the same concentrations as used in bovine [23,24] and porcine [25,26] IVM.

Techniques: