Journal: Regenerative Therapy

Article Title: Wharton's jelly mesenchymal stem cell-secretome enhances skin rejuvenation via ApoA4 and SERPINH1

doi: 10.1016/j.reth.2026.101071

Figure Lengend Snippet: Comparative in-vitro analysis of skin-rejuvenation activities of MSC secretomes. Secretomes from WJ-MSCs, AD-MSCs, and BM-MSCs were compared across multiple functional assays. Unless otherwise stated, the negative control (N.C.) was Vehicle-CM (α-MEM + 5 % human platelet lysate incubated cell-free for 48 h and processed identically). (A) HS68 fibroblast proliferation by CCK-8, normalized to N.C. (=100 %); n = 5. (B) HaCaT keratinocyte scratch-wound closure at 0 h and 18 h; n = 4. (C) Type I procollagen secretion (ELISA) in HS68 fibroblasts; n = 5. (D) ECM-related gene expression ( COL1A1, COL3A1 ) by qPCR ( GAPDH -normalized) in HS68 fibroblasts; n = 5. (E) Suppression of pro-inflammatory transcripts (IL-6/COX-2/IL-1β/TNF-α or equivalent cytokine readouts) in LPS-stimulated RAW 264.7 macrophages; n = 3. (F) HUVEC tube-formation assay (6 h); tube number and total length; n = 5. VEGF (100 ng/mL) served as positive control. (G) Intracellular ROS (DCF-DA) after H 2 O 2 challenge; secretome treatments versus N.C.; n = 3. Ascorbic acid (Vit. C) served as antioxidant control. (H) Total antioxidant capacity (reported as Trolox equivalents, nmol/μL); n = 5. Vit. C served as positive control. Data are mean ± SEM from independent experiments; statistics by one-way ANOVA with Tukey's post hoc test (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). Abbreviations: N.C., negative control (Vehicle-CM); AD, Adipose tissue-derived mesenchymal stem cells secretome; BM, bone marrow-derived mesenchymal stem cells secretome; WJ, Wharton's jelly-derived mesenchymal stem cells secretome; LPS, lipopolysaccharide; VEGF, vascular endothelial growth factor; DCF-DA, 2′,7′-dichlorofluorescein diacetate.

Article Snippet: HS68 fibroblasts (6 × 10 4 cells/well; ATCC, USA) were seeded in 6-well plates (Corning, USA) overnight and treated with WJ-MSC secretome or recombinant ApoA4/SERPINH1 (R&D Systems, USA) for 24 h. Total RNA was extracted with QIAzol (QIAGEN, Germany) and reverse transcribed using SuperScript III (Invitrogen, USA) COL1A1 and COL3A1 mRNA levels were measured by qPCR (QuantStudio 6 Flex; Thermo Fisher Scientific, USA) with SYBRTM Green Master Mix (Applied Biosystems, USA).

Techniques: In Vitro, Functional Assay, Negative Control, Incubation, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Gene Expression, HUVEC Tube Formation Assay, Positive Control, Control, Derivative Assay

Journal: Regenerative Therapy

Article Title: Wharton's jelly mesenchymal stem cell-secretome enhances skin rejuvenation via ApoA4 and SERPINH1

doi: 10.1016/j.reth.2026.101071

Figure Lengend Snippet: SERPINH1 in vitro functions relevant to skin rejuvenation. Baseline control: serum-free assay medium (SFM); Vehicle-CM not used. (A) HS68 fibroblast proliferation (CCK-8) after recombinant SERPINH1 treatment. (B) Immunoblot confirming increased cell-associated SERPINH1 after treatment. (C) HaCaT scratch-wound closure at 18 h; EGF (20 ng/mL) as positive control. (D) Immunoblot of ECM proteins (COL1A1, fibronectin). (E) Immunoblot of hydration-related proteins (HAS2, AQP3); retinoic acid as comparator (positive control). (F) siRNA-mediated SERPINH1 knockdown impairs HaCaT migration. (G) Knockdown reduces TGF-β, TGFβR1, p -Smad2/3, COL1A1, COL3A1, and fibronectin, consistent with dampened TGF-β/Smad signaling and ECM remodeling. Data are mean ± SEM; one-way ANOVA with Tukey's post hoc test (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001).

Article Snippet: HS68 fibroblasts (6 × 10 4 cells/well; ATCC, USA) were seeded in 6-well plates (Corning, USA) overnight and treated with WJ-MSC secretome or recombinant ApoA4/SERPINH1 (R&D Systems, USA) for 24 h. Total RNA was extracted with QIAzol (QIAGEN, Germany) and reverse transcribed using SuperScript III (Invitrogen, USA) COL1A1 and COL3A1 mRNA levels were measured by qPCR (QuantStudio 6 Flex; Thermo Fisher Scientific, USA) with SYBRTM Green Master Mix (Applied Biosystems, USA).

Techniques: In Vitro, Control, CCK-8 Assay, Recombinant, Western Blot, Positive Control, Knockdown, Migration

Journal: Regenerative Therapy

Article Title: Wharton's jelly mesenchymal stem cell-secretome enhances skin rejuvenation via ApoA4 and SERPINH1

doi: 10.1016/j.reth.2026.101071

Figure Lengend Snippet: Comparative in-vitro analysis of skin-rejuvenation activities of MSC secretomes. Secretomes from WJ-MSCs, AD-MSCs, and BM-MSCs were compared across multiple functional assays. Unless otherwise stated, the negative control (N.C.) was Vehicle-CM (α-MEM + 5 % human platelet lysate incubated cell-free for 48 h and processed identically). (A) HS68 fibroblast proliferation by CCK-8, normalized to N.C. (=100 %); n = 5. (B) HaCaT keratinocyte scratch-wound closure at 0 h and 18 h; n = 4. (C) Type I procollagen secretion (ELISA) in HS68 fibroblasts; n = 5. (D) ECM-related gene expression ( COL1A1, COL3A1 ) by qPCR ( GAPDH -normalized) in HS68 fibroblasts; n = 5. (E) Suppression of pro-inflammatory transcripts (IL-6/COX-2/IL-1β/TNF-α or equivalent cytokine readouts) in LPS-stimulated RAW 264.7 macrophages; n = 3. (F) HUVEC tube-formation assay (6 h); tube number and total length; n = 5. VEGF (100 ng/mL) served as positive control. (G) Intracellular ROS (DCF-DA) after H 2 O 2 challenge; secretome treatments versus N.C.; n = 3. Ascorbic acid (Vit. C) served as antioxidant control. (H) Total antioxidant capacity (reported as Trolox equivalents, nmol/μL); n = 5. Vit. C served as positive control. Data are mean ± SEM from independent experiments; statistics by one-way ANOVA with Tukey's post hoc test (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). Abbreviations: N.C., negative control (Vehicle-CM); AD, Adipose tissue-derived mesenchymal stem cells secretome; BM, bone marrow-derived mesenchymal stem cells secretome; WJ, Wharton's jelly-derived mesenchymal stem cells secretome; LPS, lipopolysaccharide; VEGF, vascular endothelial growth factor; DCF-DA, 2′,7′-dichlorofluorescein diacetate.

Article Snippet: HaCaT keratinocytes (AddexBio, San Diego, CA, USA; T0020001), HS68 human dermal fibroblasts (ATCC, Manassas, VA, USA; CRL-1635), and Raw264.7 murine macrophages (KCLB, Seoul, Korea; 40071) were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10 % fetal bovine serum (FBS; Gibco) and 1 % penicillin-streptomycin (Gibco) at 37 °C in a humidified 5 % CO 2 incubator.

Techniques: In Vitro, Functional Assay, Negative Control, Incubation, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Gene Expression, HUVEC Tube Formation Assay, Positive Control, Control, Derivative Assay

Journal: Regenerative Therapy

Article Title: Wharton's jelly mesenchymal stem cell-secretome enhances skin rejuvenation via ApoA4 and SERPINH1

doi: 10.1016/j.reth.2026.101071

Figure Lengend Snippet: SERPINH1 in vitro functions relevant to skin rejuvenation. Baseline control: serum-free assay medium (SFM); Vehicle-CM not used. (A) HS68 fibroblast proliferation (CCK-8) after recombinant SERPINH1 treatment. (B) Immunoblot confirming increased cell-associated SERPINH1 after treatment. (C) HaCaT scratch-wound closure at 18 h; EGF (20 ng/mL) as positive control. (D) Immunoblot of ECM proteins (COL1A1, fibronectin). (E) Immunoblot of hydration-related proteins (HAS2, AQP3); retinoic acid as comparator (positive control). (F) siRNA-mediated SERPINH1 knockdown impairs HaCaT migration. (G) Knockdown reduces TGF-β, TGFβR1, p -Smad2/3, COL1A1, COL3A1, and fibronectin, consistent with dampened TGF-β/Smad signaling and ECM remodeling. Data are mean ± SEM; one-way ANOVA with Tukey's post hoc test (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001).

Article Snippet: HaCaT keratinocytes (AddexBio, San Diego, CA, USA; T0020001), HS68 human dermal fibroblasts (ATCC, Manassas, VA, USA; CRL-1635), and Raw264.7 murine macrophages (KCLB, Seoul, Korea; 40071) were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco, Grand Island, NY, USA) supplemented with 10 % fetal bovine serum (FBS; Gibco) and 1 % penicillin-streptomycin (Gibco) at 37 °C in a humidified 5 % CO 2 incubator.

Techniques: In Vitro, Control, CCK-8 Assay, Recombinant, Western Blot, Positive Control, Knockdown, Migration