Journal: Bioactive Materials

Article Title: Multifactorial bioengineering in the tendon context to maintain tenocyte phenotype and to direct dermal fibroblasts towards tenogenic lineage

doi: 10.1016/j.bioactmat.2025.11.008

Figure Lengend Snippet: Immunofluorescence intensity analysis for COL I, III, IV, V, VI, COL I/COL III ratio, TNMD and aSMA in hTCs and hDFs after 4, 7 and 10 days of culture under PLLA, +TGFB2, +MMC and +MMC + TGFB2 conditions. (∗p < 0.05 vs respective PLLA group at the same time point; ‡p < 0.05 vs respective + TGFB2 group at the same time point; #p < 0.05 vs. respective + MMC group at the same time point; §p < 0.05 between cell types at the same time point and condition; &p < 0.05 vs. respective condition at day 4). N = 3.

Article Snippet: Cryopreserved hDFs (PCS-201-012) were purchased from the American Type Culture Collection (ATCC, United Kingdom).

Techniques: Immunofluorescence

Journal: Bioactive Materials

Article Title: Multifactorial bioengineering in the tendon context to maintain tenocyte phenotype and to direct dermal fibroblasts towards tenogenic lineage

doi: 10.1016/j.bioactmat.2025.11.008

Figure Lengend Snippet: Principal component analysis ( A ) of the quantitative proteomic data obtained from hTCs and hDFs after 10 days of culture under TCP, PLLA, +TGFB2, +MMC and +MMC + TGFB2 conditions, as well as from TT samples. Venn diagram representing the top 100 protein hits in TT group vs. hTCs ( B ) and hDFs ( C ) cultured under PS, PLLA, +TGFB2, +MMC and +MMC + TGFB2 conditions. Gene ontology analysis ( D ) of the top 100 protein hits in TT group vs. hTCs and hDFs cultured under PS, PLLA, +TGFB2, +MMC and +MMC + TGFB2 conditions. N = 3.

Article Snippet: Cryopreserved hDFs (PCS-201-012) were purchased from the American Type Culture Collection (ATCC, United Kingdom).

Techniques: Cell Culture

Journal: Bioactive Materials

Article Title: Multifactorial bioengineering in the tendon context to maintain tenocyte phenotype and to direct dermal fibroblasts towards tenogenic lineage

doi: 10.1016/j.bioactmat.2025.11.008

Figure Lengend Snippet: Gene set enrichment analysis of the quantitative proteomic profile of hDFs cultured for 10 days under PLLA ( A ), +TGFB2 ( B ), +MMC ( C ), and +MMC + TGFB2 ( D ) conditions. All contrasts were performed against hDFs cultured for 10 days under TCP conditions. A gene set containing all protein hits in TT group vs hDFs TCP group (tendon signature) was used for all analyses. Venn diagram summarizing the different core enrichment proteins identified in PLLA, +TGFB2, +MMC and +MMC + TGFB2 groups by means of gene set enrichment analysis ( E ). GO analysis of the core enrichment proteins identified in +MMC + TGFB2 group by means of gene set enrichment analysis ( F ). (NES: normalised enrichment score). N = 3.

Article Snippet: Cryopreserved hDFs (PCS-201-012) were purchased from the American Type Culture Collection (ATCC, United Kingdom).

Techniques: Cell Culture

Journal: Bioactive Materials

Article Title: Multifactorial bioengineering in the tendon context to maintain tenocyte phenotype and to direct dermal fibroblasts towards tenogenic lineage

doi: 10.1016/j.bioactmat.2025.11.008

Figure Lengend Snippet: Heatmap displaying the quantitative matrisome of hTCs and hDFs cultured under PLLA, +TGFB2, +MMC and +MMC + TGFB2 conditions for 10 days. Hierarchical clustering was applied to all samples. The heatmap displays the z-scores for every matrisome protein identified, being these grouped into the different matrisome categories. Both hTCs and hDFs clustered first in relation to TGFB2 treatment (alone or in combination with MMC), and ultimately in relation with the experimental conditions to which these were subjected. The + MMC + TGFB2 experimental condition exerted the highest changes into the quantitative matrisome of hTCs and hDFs, increasing ECM protein levels throughout all the categories of the matrisome. N = 3.

Article Snippet: Cryopreserved hDFs (PCS-201-012) were purchased from the American Type Culture Collection (ATCC, United Kingdom).

Techniques: Cell Culture

Journal: Stem Cell Research & Therapy

Article Title: Immortalized human hair follicle-derived mesenchymal-like stromal cells for the long-term production of scalable Immunomodulatory and regenerative secretome

doi: 10.1186/s13287-025-04775-8

Figure Lengend Snippet: Regenerative effect of iHF-MSCs. A – B Effect of iHF-MSC-derived unlicensed conditioned media on the proliferation of HaCaTs. Scale bars = 200 μm. C – D Effect of iHF-MSC-derived unlicensed conditioned media on the proliferation of HDFs. Scale bars = 200 μm. E In vitro wound closure assay in HaCaTs. Scale bars = 200 μm. F In vitro wound closure assay in HDFs. Scale bars = 200 μm. G HUVEC-mediated tube formation assay. Scale bars = 200 μm. Data are presented as mean ± SD. N = 3 independent experiments, with 5 replicates per group. Statistical significance: γ p < 0.05, γγ p < 0.01, γγγ p < 0.001 compared to the negative control; * p < 0.05, ** p < 0.01 and *** p < 0.001 compared to HF-MSCs ## p < 0.01 compared to C18; $ p < 0.05, $$ p < 0.01 and $$$ p < 0.001 compared to the positive control. One-way ANOVA with the Bonferroni post-hoc test in B , D , E – II , F-II , G-II , G-III; Student’s t-test in G-IV . HF-MSCs: hair follicle-derived mesenchymal stromal cells. C18: clone 18. C26: clone 26. HaCaTs: HaCaT keratinocytes. HDFs: adult dermal fibroblasts. HUVECs: human umbilical vein endothelial cells

Article Snippet: The cells used in this study comprise adult human dermal fibroblasts (HDFs) (ATCC, PCS-201-012), human umbilical vein endothelial cells (HUVECs) (Lonza, C2519A), HaCaT keratinocytes (DKFZ, Germany), HF-MSCs, iHF-MSCs and PBMCs.

Techniques: Derivative Assay, In Vitro, Wound Closure Assay, Tube Formation Assay, Negative Control, Positive Control

Journal: Stem Cell Research & Therapy

Article Title: Immortalized human hair follicle-derived mesenchymal-like stromal cells for the long-term production of scalable Immunomodulatory and regenerative secretome

doi: 10.1186/s13287-025-04775-8

Figure Lengend Snippet: iHF-MSC protection against oxidative stress. A Neutralization effect and B protective effect of iHF-MSC-derived unlicensed conditioned media over HaCaTs. C Neutralization effect and D protective effect of iHF-MSC-derived unlicensed conditioned media over HDFs. E – I DPPH scavenging capacity of iHF-MSCs. E – II DPPH scavenging percentage at 0 and 4 h. F Survival ratio of HaCaTs exposed to hyperglycemia upon treatment with iHF-MSC-derived unlicensed conditioned media. G Survival ratio of HDFs exposed to hyperglycemia upon treatment with iHF-MSC-derived unlicensed conditioned media. Data are presented as mean ± SD. N = 3 independent experiments, with 4 replicates per group. Statistical significance: γ p < 0.05, γγ p < 0.01, γγγ p < 0.001 compared to the negative control; * p < 0.05, ** p < 0.01 and *** p < 0.001 compared to HF-MSCs; ### p < 0.001 compared to C18; $ p < 0.05, $$ p < 0.01 and $$$ p < 0.001 compared to the positive control; ββ p < 0.01 and βββ p < 0.001 compared to W/o H 2 O 2 or W/o Glucose groups. One-way ANOVA with the Bonferroni post-hoc test in A-G . HF-MSCs: hair follicle-derived mesenchymal stromal cells. C18: clone 18. C26: clone 26. W/o H2O2: without hydrogen peroxide. W/o Glucose: without glucose

Article Snippet: The cells used in this study comprise adult human dermal fibroblasts (HDFs) (ATCC, PCS-201-012), human umbilical vein endothelial cells (HUVECs) (Lonza, C2519A), HaCaT keratinocytes (DKFZ, Germany), HF-MSCs, iHF-MSCs and PBMCs.

Techniques: Neutralization, Derivative Assay, Negative Control, Positive Control

Journal: Stem Cell Research & Therapy

Article Title: Immortalized human hair follicle-derived mesenchymal-like stromal cells for the long-term production of scalable Immunomodulatory and regenerative secretome

doi: 10.1186/s13287-025-04775-8

Figure Lengend Snippet: Regenerative effect of iHF-MSCs. A – B Effect of iHF-MSC-derived unlicensed conditioned media on the proliferation of HaCaTs. Scale bars = 200 μm. C – D Effect of iHF-MSC-derived unlicensed conditioned media on the proliferation of HDFs. Scale bars = 200 μm. E In vitro wound closure assay in HaCaTs. Scale bars = 200 μm. F In vitro wound closure assay in HDFs. Scale bars = 200 μm. G HUVEC-mediated tube formation assay. Scale bars = 200 μm. Data are presented as mean ± SD. N = 3 independent experiments, with 5 replicates per group. Statistical significance: γ p < 0.05, γγ p < 0.01, γγγ p < 0.001 compared to the negative control; * p < 0.05, ** p < 0.01 and *** p < 0.001 compared to HF-MSCs ## p < 0.01 compared to C18; $ p < 0.05, $$ p < 0.01 and $$$ p < 0.001 compared to the positive control. One-way ANOVA with the Bonferroni post-hoc test in B , D , E – II , F-II , G-II , G-III; Student’s t-test in G-IV . HF-MSCs: hair follicle-derived mesenchymal stromal cells. C18: clone 18. C26: clone 26. HaCaTs: HaCaT keratinocytes. HDFs: adult dermal fibroblasts. HUVECs: human umbilical vein endothelial cells

Article Snippet: Commercial cell lines, including HDFs (ATCC, PCS-201-012), HUVECs (Lonza, C2519A), and HaCaTs (DKFZ, Germany), were acquired from certified repositories and used in accordance with institutional biosafety and ethical guidelines.

Techniques: Derivative Assay, In Vitro, Wound Closure Assay, Tube Formation Assay, Negative Control, Positive Control

Journal: Stem Cell Research & Therapy

Article Title: Immortalized human hair follicle-derived mesenchymal-like stromal cells for the long-term production of scalable Immunomodulatory and regenerative secretome

doi: 10.1186/s13287-025-04775-8

Figure Lengend Snippet: iHF-MSC protection against oxidative stress. A Neutralization effect and B protective effect of iHF-MSC-derived unlicensed conditioned media over HaCaTs. C Neutralization effect and D protective effect of iHF-MSC-derived unlicensed conditioned media over HDFs. E – I DPPH scavenging capacity of iHF-MSCs. E – II DPPH scavenging percentage at 0 and 4 h. F Survival ratio of HaCaTs exposed to hyperglycemia upon treatment with iHF-MSC-derived unlicensed conditioned media. G Survival ratio of HDFs exposed to hyperglycemia upon treatment with iHF-MSC-derived unlicensed conditioned media. Data are presented as mean ± SD. N = 3 independent experiments, with 4 replicates per group. Statistical significance: γ p < 0.05, γγ p < 0.01, γγγ p < 0.001 compared to the negative control; * p < 0.05, ** p < 0.01 and *** p < 0.001 compared to HF-MSCs; ### p < 0.001 compared to C18; $ p < 0.05, $$ p < 0.01 and $$$ p < 0.001 compared to the positive control; ββ p < 0.01 and βββ p < 0.001 compared to W/o H 2 O 2 or W/o Glucose groups. One-way ANOVA with the Bonferroni post-hoc test in A-G . HF-MSCs: hair follicle-derived mesenchymal stromal cells. C18: clone 18. C26: clone 26. W/o H2O2: without hydrogen peroxide. W/o Glucose: without glucose

Article Snippet: Commercial cell lines, including HDFs (ATCC, PCS-201-012), HUVECs (Lonza, C2519A), and HaCaTs (DKFZ, Germany), were acquired from certified repositories and used in accordance with institutional biosafety and ethical guidelines.

Techniques: Neutralization, Derivative Assay, Negative Control, Positive Control