Journal: International Journal of Biological Sciences

Article Title: Human umbilical cord mesenchymal stem cell delivery of mitochondria to melanocytes enhances skin repigmentation efficacy in autologous epidermal cell suspension transplantation through the TNFAIP2-TNT system

doi: 10.7150/ijbs.128719

Figure Lengend Snippet: Co-culture with hUCMSCs enhances the function of ECs. (A) Diagram of direct co-culture (dir. co) and indirect co-culture (ind. co) process. (B) Observation of ECs alone and ECs+MSCs (dir. co) by light microscope. Scale bars: 100 μm. (C) Flow cytometry of MCs in ECs comparison of ECs alone and ECs+MSCs (dir. co). (D) Statistical analysis of MCs ratio in ECs and ECs+MSCs at week 1, week 2 and week 3. (E) Counted and calculated the total number of the absolute number of MCs, then compared it with the baseline number to determine the proliferation rate of MCs. (F-G) Proliferative ability of MCs and KCs (ind. co) at week 2 by CCK-8. (H-I) Migration ability of MC and KCs compared with hUCMSCs co-cultured (ind. co) at 72 h. Scale bars: 200 μm. (J) Transwell assay on ECs and ECs+MSCs (dir. co) with masson-fontana to identify MCs. Scale bars: 100 μm. (K-L) Immunofluorescent staining of MCs alone and MCs + hUCMSCs at week 2 suggesting the differentiation situation and adhesion ability of MCs. Scale bars: 30 μm. (M) Digested and collected the cells after culturing ECs alone and ECs+MSCs for three weeks. (N) Tyrosinase activity of ECs alone and ECs + hUCMSCs after co-cultured for 1 w, 2 w and 3 w. (O) The ratio of tyrosinase activity measured in 1×10 5 cells before and after co-cultured for one week. (P) Melanin content of 1×10 5 ECs alone and ECs + hUCMSCs after co-cultured for 1 w, 2 w and 3 w. (Q) The ratio of melanin content measured in 1×10 5 cells before and after co-cultured for one week. Data show mean ± S.D, and each experiment was conducted with a minimum of three replicates. * P ≤0.05, ** P ≤0.01, **** P ≤0.0001, by two-tailed Student's t test.

Article Snippet: Subsequently, hUCMSCs were isolated using CD105 MicroBeads for humans (130-051-201, Miltenyi Biotec, Germany), and the cells obtained through negative selection were identified as MCs.

Techniques: Co-Culture Assay, Light Microscopy, Flow Cytometry, Comparison, CCK-8 Assay, Migration, Cell Culture, Transwell Assay, Staining, Activity Assay, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: Human umbilical cord mesenchymal stem cell delivery of mitochondria to melanocytes enhances skin repigmentation efficacy in autologous epidermal cell suspension transplantation through the TNFAIP2-TNT system

doi: 10.7150/ijbs.128719

Figure Lengend Snippet: Co-transplantation of hUCMSCs enhances the function of ECs. (A) Diagram of transplanting the prepared AECS onto the polished back of nude mice process. (B) After transplantation for 21 days, the transplantation sites were observed under white light, wood light, and skin microscope. Scale bars: 500 μm. (C) Changes in skin pigmentation were assessed spectrophotometrically and ΔL values were quantified before and after animal transplantation. (D) After transplantation, the source of ECs in the corresponding sites were represented by human reactive K10 (hu-K10) and human/mouse reactive K5 (hu/ms-K5). Scale bars: 100 μm. (E) Light microscopy of skin specimens stained with H&E, Melan-A immunohistochemistry, and Fontana-Masson to compare the residing location of MCs and melanin content. Scale bars: 100 μm. (F-H) Immunofluorescence staining: Skin specimens stained with Melan-A, TRP-1, DCT and TUNEL apoptosis assay to detect the number, mature proportion, and apoptosis of MCs. Scale bars: 100 μm. (I-K) Semi-quantitative analysis of the fluorescence intensity. Data show mean ± S.D, and each experiment was conducted with a minimum of three replicates. * P ≤0.05, ** P ≤0.01, *** P ≤0.001, **** P ≤0.0001, ns indicates no significant difference (n=3; one-way ANOVA with Bonferroni's post-test analysis).

Article Snippet: Subsequently, hUCMSCs were isolated using CD105 MicroBeads for humans (130-051-201, Miltenyi Biotec, Germany), and the cells obtained through negative selection were identified as MCs.

Techniques: Transplantation Assay, Microscopy, Light Microscopy, Staining, Immunohistochemistry, Immunofluorescence, TUNEL Assay, Apoptosis Assay, Fluorescence

Journal: International Journal of Biological Sciences

Article Title: Human umbilical cord mesenchymal stem cell delivery of mitochondria to melanocytes enhances skin repigmentation efficacy in autologous epidermal cell suspension transplantation through the TNFAIP2-TNT system

doi: 10.7150/ijbs.128719

Figure Lengend Snippet: hUCMSCs co-culture enhances cytoskeleton and cell-cell connection pathway. (A-B) Histogram and Volcano plot depicted differential expressing genes of MC samples: MCs co-cultured with hUCMSCs (co-MCs) versus MCs alone (MCs). (C) Differential gene analysis heatmap: co-MCs versus MCs. (D) Bulk RNA-seq of MC samples: KEGG analysis (positive enrichment) for co-MCs versus MCs. (E) GO analysis (positive enrichment) for co-MCs versus MCs. (F) Captured the connections between cells with SEM. Scale bars: 500nm, 200nm. (G) Immunofluorescent staining cell-cell connection between hUCMSCs and MCs was showed using TNTs compositions, Tubulin and F-actin. Scale bars: 30 μm.

Article Snippet: Subsequently, hUCMSCs were isolated using CD105 MicroBeads for humans (130-051-201, Miltenyi Biotec, Germany), and the cells obtained through negative selection were identified as MCs.

Techniques: Co-Culture Assay, Expressing, Cell Culture, RNA Sequencing, Staining

Journal: International Journal of Biological Sciences

Article Title: Human umbilical cord mesenchymal stem cell delivery of mitochondria to melanocytes enhances skin repigmentation efficacy in autologous epidermal cell suspension transplantation through the TNFAIP2-TNT system

doi: 10.7150/ijbs.128719

Figure Lengend Snippet: hUCMSCs enhance MCs function by forming TNTs and mitochondrial transfer. (A-B) Immunofluorescent staining revealed the transfer of mitochondria between the two cells, using MTR-hUCMSCs and MTG-MCs. Scale bars: 30 μm. (C) Flow cytometry to statistically analyze the proportion of MCs that have received mitochondria from hUCMSCs. (D) Schematic diagram of the composition of TNTs and the inhibitors of its corresponding components: Nocodazole (Noc, tubulin inhibitor), Cytochalasin D (CyD, F-actin inhibitor). (E) Observed the relationship between ATP levels of MCs and the formation of TNTs. (F-G) JC-1 assay for measuring mitochondria depolarization by immunofluorescent staining and microplate reader. Scale bars: 30 μm. (H) Apoptosis level of MCs before and after blocking the formation of TNTs. (I) Blocked the formation of TNTs and observed the oxidative stress levels by flow cytometry. Using Rosup as a positive control, calculated the relative fluorescence intensity of each group. (J) Confocal microscope detected the intracellular ROS level, and the green fluorescence intensity was summarized in the bar graph. (K) Microplate reader detected ROS level and calculated the relative ROS level using MCs as control. (L) Migration ability of MCs before and after blocking the formation of TNTs by transwell assay. Scale bars: 100 μm. (M-N) Observed the relationship between proliferative ability of MCs and the formation of TNTs: light microscope, CCK-8 assay. Scale bars: 150 μm. (O-P) Melanin content and the ability to synthesize melanin of MCs with or without TNTs blocking by NaOH cracking method and tyrosinase activity. Data show mean ± S.D, and each experiment was conducted with a minimum of three replicates. * P ≤0.05, ** P ≤0.01, *** P ≤0.001, **** P ≤0.0001, by two-tailed Student's t test and one-way ANOVA with Bonferroni's post-test analysis.

Article Snippet: Subsequently, hUCMSCs were isolated using CD105 MicroBeads for humans (130-051-201, Miltenyi Biotec, Germany), and the cells obtained through negative selection were identified as MCs.

Techniques: Staining, Flow Cytometry, Blocking Assay, Positive Control, Fluorescence, Microscopy, Control, Migration, Transwell Assay, Light Microscopy, CCK-8 Assay, Activity Assay, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: Human umbilical cord mesenchymal stem cell delivery of mitochondria to melanocytes enhances skin repigmentation efficacy in autologous epidermal cell suspension transplantation through the TNFAIP2-TNT system

doi: 10.7150/ijbs.128719

Figure Lengend Snippet: TNTs mediate the enhancement of MCs functions by hUCMSCs. (A) After transplantation, the source of ECs in the corresponding sites were represented by hu-K10 and hu/ms-K5. Scale bars: 100 μm. (B) After transplantation for 21 days, the transplantation sites were observed under white light, wood light, and skin microscope. Scale bars: 500 μm. (C) Changes in skin pigmentation were assessed spectrophotometrically and ΔL values were quantified before and after animal transplantation. (D) Light microscopy of skin specimens stained with H&E, Melan-A immunohistochemistry, and Fontana-Masson to compare the residing location of MCs and melanin content. Scale bars: 100 μm. (E-F) Immunofluorescence staining: melan-A, TUNEL apoptosis assay to detect the number, apoptosis condition of MCs. Scale bars: 100 μm. (G-H) Semi-quantitative analysis of the fluorescence intensity. (I-J) Immunofluorescence staining of TRP-1 and DCT to detect mature proportion of MCs. Scale bars: 100 μm. Data show mean ± S.D, and each experiment was conducted with a minimum of three replicates. * P ≤0.05, ** P ≤0.01, *** P ≤0.001, **** P ≤0.0001, ns indicates no significant difference (n=3; two-tailed Student's t test).

Article Snippet: Subsequently, hUCMSCs were isolated using CD105 MicroBeads for humans (130-051-201, Miltenyi Biotec, Germany), and the cells obtained through negative selection were identified as MCs.

Techniques: Transplantation Assay, Microscopy, Light Microscopy, Staining, Immunohistochemistry, Immunofluorescence, TUNEL Assay, Apoptosis Assay, Fluorescence, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: Human umbilical cord mesenchymal stem cell delivery of mitochondria to melanocytes enhances skin repigmentation efficacy in autologous epidermal cell suspension transplantation through the TNFAIP2-TNT system

doi: 10.7150/ijbs.128719

Figure Lengend Snippet: TNFAIP2 knockdown inhibits mitochondrial transfer and the function of MCs. (A) Volcano plot depicted the upregulation of TNTs regulating gene in co-MCs group. (B-C) Verified the target genes at the RNA and protein levels using qPCR and WB. (D-E) Immunofluorescence staining, and flow cytometry were used to statistically analyze the proportion of TNFAIP2-KD MCs that received mitochondria from hUCMSCs. Scale bars: 30 μm. (F) ATP levels in TNFAIP2-KD MCs co-cultured with hUCMSCs. (G-H) The JC-1 assay was used to measure mitochondrial depolarization through immunofluorescent staining and analysis with a microplate reader. Scale bars: 30 μm. (I-J) The oxidative stress levels in TNFAIP2-KD MCs by immunofluorescent staining and flow cytometry. Scale bars: 30 μm. (K) The apoptosis rates in TNFAIP2-KD MCs. (L) Assessment of melanin synthesis capability in TNFAIP2-KD MCs via Tyrosinase Activity Assay. (M-N) The proliferation status of TNFAIP2-KD MCs observed under a light microscope and assessed by CCK-8 assay. Scale bars: 100 μm. (O) The migration of TNFAIP2-KD MCs by transwell assay. Scale bars: 100 μm. (P) NaOH assay measuring the content of melanin changes in TNFAIP2-KD MCs. Data show mean ± S.D, and each experiment was conducted with a minimum of three replicates. * P ≤0.05, ** P ≤0.01, *** P ≤0.001, **** P ≤0.0001, by two-tailed Student's t test.

Article Snippet: Subsequently, hUCMSCs were isolated using CD105 MicroBeads for humans (130-051-201, Miltenyi Biotec, Germany), and the cells obtained through negative selection were identified as MCs.

Techniques: Knockdown, Immunofluorescence, Staining, Flow Cytometry, Cell Culture, Activity Assay, Light Microscopy, CCK-8 Assay, Migration, Transwell Assay, Two Tailed Test

Journal: International Journal of Biological Sciences

Article Title: Human umbilical cord mesenchymal stem cell delivery of mitochondria to melanocytes enhances skin repigmentation efficacy in autologous epidermal cell suspension transplantation through the TNFAIP2-TNT system

doi: 10.7150/ijbs.128719

Figure Lengend Snippet: TNFAIP2 overexpression enhances mitochondria transfer and the function of MCs. (A-B) After treatment with TNFAIP2-OE plasmids, immunofluorescence staining, and flow cytometry were used to statistically analyze the proportion of MCs that had received mitochondria from hUCMSCs. Scale bars: 30 μm. (C) ATP levels in TNFAIP2-OE MCs co-cultured with hUCMSCs. (D-E) The JC-1 assay for measuring mitochondrial depolarization in TNFAIP2-overexpressing MCs co-cultured with hUCMSCs using immunofluorescent staining and a microplate reader. Scale bars: 30 μm. (F-G) The oxidative stress levels in TNFAIP2-OE MCs co-cultured with hUCMSCs by immunofluorescent staining and flow cytometry. Scale bars: 30 μm. (H) Apoptosis level in TNFAIP2-OE MCs after co-cultured with hUCMSCs. (I) Assessment of melanin synthesis capability in TNFAIP2-OE MCs via Tyrosinase Activity Assay. (J-K) After treatment with TNFAIP2-OE plasmids, the proliferation status of MCs was observed under a light microscope and assessed using a CCK-8 assay. Scale bars: 150 μm. (L) After treatment with TNFAIP2-OE plasmids, the migration of MCs was assessed using a transwell assay. Scale bars: 100 μm. (M) NaOH assay measuring the content of melanin changes in TNFAIP2-KD MCs. Data show mean ± S.D, and each experiment was conducted with a minimum of three replicates. * P ≤0.05, ** P ≤0.01, **** P ≤0.0001, by two-tailed Student's t test.

Article Snippet: Subsequently, hUCMSCs were isolated using CD105 MicroBeads for humans (130-051-201, Miltenyi Biotec, Germany), and the cells obtained through negative selection were identified as MCs.

Techniques: Over Expression, Immunofluorescence, Staining, Flow Cytometry, Cell Culture, Activity Assay, Light Microscopy, CCK-8 Assay, Migration, Transwell Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Human umbilical cord mesenchymal stem cell–derived exosomes are associated with changes in renal injury markers, gut microbiota composition, and inflammatory signaling in IgA nephropathy

doi: 10.3389/fimmu.2026.1854005

Figure Lengend Snippet: hUCMSC-Exos modulate gut microbiota composition in IgAN-like mice. (A) Principal coordinate analysis (PCoA) based on Bray-Curtis distance showing microbial community structure among Control (n=5), IgAN (n=5), and IgAN+Exos (n=5) groups. PERMANOVA: R² = 0.4102, P = 0.001. (B) Genera previously associated with differentially enriched taxa in other studies are highlighted. (C) Spearman correlation analysis between representative genera and renal function/inflammatory parameters (Scr, UPCR, BUN, and serum NLRP3). Correlations are exploratory; uncorrected P values are shown. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 (Kruskal-Wallis test with Dunn’s post-hoc for B; Spearman correlation for C). Details are provided in Methods.

Article Snippet: When passage-5 hUCMSCs reached approximately 70% confluence, the medium was replaced with exosome-depleted medium, and cells were cultured for an additional 48h. hUCMSC-Exos were isolated by differential ultracentrifugation according to MISEV2023 guidelines ( ).

Techniques: Control

Journal: Frontiers in Immunology

Article Title: Human umbilical cord mesenchymal stem cell–derived exosomes are associated with changes in renal injury markers, gut microbiota composition, and inflammatory signaling in IgA nephropathy

doi: 10.3389/fimmu.2026.1854005

Figure Lengend Snippet: hUCMSC-Exos are associated with reduced renal dysfunction and histopathological injury in IgAN-like mice. (A) Blood urea nitrogen (BUN). (B) Serum creatinine (Scr). (C) Serum NLRP3 levels. (D) Urinary protein-to-creatinine ratio (UPCR). (E) Representative histological images are shown; quantitative histopathological scoring could not be performed due to limited remaining tissue availability. (F) Immunofluorescence analysis of glomerular IgA deposition. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 (one-way ANOVA with Tukey’s post hoc test for normally distributed data; otherwise Kruskal–Wallis test with Dunn’s correction). Each dot represents one animal.

Article Snippet: When passage-5 hUCMSCs reached approximately 70% confluence, the medium was replaced with exosome-depleted medium, and cells were cultured for an additional 48h. hUCMSC-Exos were isolated by differential ultracentrifugation according to MISEV2023 guidelines ( ).

Techniques: Immunofluorescence

Journal: Frontiers in Immunology

Article Title: Human umbilical cord mesenchymal stem cell–derived exosomes are associated with changes in renal injury markers, gut microbiota composition, and inflammatory signaling in IgA nephropathy

doi: 10.3389/fimmu.2026.1854005

Figure Lengend Snippet: hUCMSC-Exos are associated with reduced podocyte injury and NLRP3 inflammasome-related responses in vitro . (A) Podocyte viability assessed by CCK-8 assay. (B–D) Levels of IL-1β, IL-18, and NLRP3 in culture supernatants measured by ELISA. (E) NLRP3 mRNA expression analyzed by RT-qPCR. (F) Representative Western blot analysis of NLRP3 protein expression (n=3 independent experiments). All in vitro experiments were performed with three independent biological replicates, each with three technical replicates. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 (one-way ANOVA with Tukey’s post-hoc test).

Article Snippet: When passage-5 hUCMSCs reached approximately 70% confluence, the medium was replaced with exosome-depleted medium, and cells were cultured for an additional 48h. hUCMSC-Exos were isolated by differential ultracentrifugation according to MISEV2023 guidelines ( ).

Techniques: In Vitro, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Immunology

Article Title: Human umbilical cord mesenchymal stem cell–derived exosomes are associated with changes in renal injury markers, gut microbiota composition, and inflammatory signaling in IgA nephropathy

doi: 10.3389/fimmu.2026.1854005

Figure Lengend Snippet: Proposed association model of hUCMSC-Exos in IgAN-like mice. Schematic illustration summarizing the associations observed in this study. hUCMSC-Exos treatment was associated with altered gut microbiota composition and reduced inflammatory-related markers in podocytes and renal tissues. Potential interactions among gut microbiota, microbial metabolites, and host inflammatory responses are presented as hypothetical associations. Solid arrows indicate experimentally observed associations, whereas dashed arrows indicate inferred or speculative relationships that were not directly tested in this study.

Article Snippet: When passage-5 hUCMSCs reached approximately 70% confluence, the medium was replaced with exosome-depleted medium, and cells were cultured for an additional 48h. hUCMSC-Exos were isolated by differential ultracentrifugation according to MISEV2023 guidelines ( ).

Techniques: