Journal: Molecular Therapy

Article Title: Phase 1 dose-escalation trial of sub-endometrial injection of human embryonic stem cells-derived immunity-and-matrix-regulatory cells to promote endometrial angiogenesis in refractory intrauterine adhesion

doi: 10.1016/j.ymthe.2025.09.035

Figure Lengend Snippet: The expression levels of angiogenetic proteins in conditional medium (A and B) Fluorescence intensity of differentially expressed proteins among three groups: ESC-CM, IMRC-CM, and co-culture-CM. ANG-1, angiopoietin-1;ANGPTL4, angiopoietin-like-4; CXCL16, chemokine ligand-16; ENA-78, epithelial neutrophil-activating protein-78; FGF, fibroblast growth factor; GRO, growth-related oncogene; HGF, hepatocyte growth factor; IL-8, interleukin-8; IP-10, interferon-gamma-inducible protein-10; MCP-1, monocyte chemoattractant protein-1; MMP-1, matrix metalloproteinases-1; TIMP-1, tissue inhibitor of metalloproteinase-1; TPO, thrombopoietin; uPAR, urokinase-type plasminogen activator receptor. The dots in the bar chart represent the number of technical replicates. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; ns, not significant. Data shown as mean ± SD.

Article Snippet: The concentrations of ANGPTL4 (catalog no.70-EK1292-96, Liankebio, China) in ESC-CM, IMRC-CM, and co-culture-CM were measured using human ELISA kits.

Techniques: Expressing, Fluorescence, Co-Culture Assay

Journal: Molecular Therapy

Article Title: Phase 1 dose-escalation trial of sub-endometrial injection of human embryonic stem cells-derived immunity-and-matrix-regulatory cells to promote endometrial angiogenesis in refractory intrauterine adhesion

doi: 10.1016/j.ymthe.2025.09.035

Figure Lengend Snippet: ANGPTL4 plays a key role in co-culture-CM-induced angiogenesis (A) The protein concentration of ANGPTL4 was measured by ELISA among ESC-CM, IMRC-CM, and co-culture-CM groups, respectively. (B and C) The mRNA expression of ANGPTL4 in ESCs (B) and IMRCs (C) was measured by quantitative real-time PCR. Co-cultured ESCs: ESC after co-culturing with IMRC; co-cultured IMRCs: IMRCs after co-cultured with ESCs. (D) The protein expression of ANGPTL4 in co-cultured ESCs was measured by Western blotting. (E) The relative expression of ANGPTL4 after siRNA-mediated knockdown of ANGPTL4 in ESCs. (F–H) Co-culture-CM-induced angiogenesis was impaired after the knockdown of ANGPTL4 in ESCs. Ctrl, control, basal medium for IMRCs; si-NC, si-ANGPTL4, siRNA of ANGPTL4; siRNA of negative control. Scale bar, 100 μm. (I–K) Angiogenesis increased in the rHuANGPTL4 group compared to that in the Ctrl group. A similar result was observed in the co-culture-CM and rHuANGPTL4 combined groups in comparison with that in co-culture-CM alone. The concentration of rHuANGPTL4 was 1 μg/mL. Scale bar, 100 μm. The dots in the bar chart represent the number of technical replicates. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, not significant. Data shown as mean ± SD.

Article Snippet: The concentrations of ANGPTL4 (catalog no.70-EK1292-96, Liankebio, China) in ESC-CM, IMRC-CM, and co-culture-CM were measured using human ELISA kits.

Techniques: Co-Culture Assay, Protein Concentration, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Cell Culture, Western Blot, Knockdown, Control, Negative Control, Comparison, Concentration Assay

Journal: Molecular Therapy

Article Title: Phase 1 dose-escalation trial of sub-endometrial injection of human embryonic stem cells-derived immunity-and-matrix-regulatory cells to promote endometrial angiogenesis in refractory intrauterine adhesion

doi: 10.1016/j.ymthe.2025.09.035

Figure Lengend Snippet: IMRCs significantly promote endometrial angiogenesis in IUA patients (A–C) The hysteroscopic image (A), H&E staining (B), and Masson staining (C) of the endometrium of patient 16 at V3 after cell injection. Scale bar, 100 µm. The hysteroscopic image (A) of patient 16 at visit V3 is also presented in A, as both figures correspond to the same patient at the same follow-up time point. (D and E) The representative images (D) and quantitative analysis (E) of immunofluorescence of CD31 and α-SMA in the normal proliferative endometrium, IUA patient endometrium, and the endometrium of patient 16 at V3. Scale bar, 50 μm. (F and G) The representative images (F) and quantitative analysis (G) of immunohistochemistry of ANGPTL4 in normal proliferative endometrium, endometrium of patient 16 at V0, and endometrium of patient 16 at V3. Scale bar, 100 µm. The dots in the bar chart represent the number of biological replicates, which is three. ∗ p < 0.05; ∗∗ p < 0.01. Data shown as mean ± SD.

Article Snippet: The concentrations of ANGPTL4 (catalog no.70-EK1292-96, Liankebio, China) in ESC-CM, IMRC-CM, and co-culture-CM were measured using human ELISA kits.

Techniques: Staining, Injection, Immunofluorescence, Immunohistochemistry