Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Colorimetric staining and quantification of ALP activity in hBM-MSCs cultured for seven days on collagen type I-coated (a) and fibronectin-coated (b) substrates. Results are expressed as mean ± SD (n=3). Data were analyzed assuming a Gaussian (normal) distribution and equal variances across groups; statistical differences were assessed using one-way ANOVA. Only p-values <0.1 are shown. Scale bar represents 200 µm.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Staining, Activity Assay, Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Fluorescence images of hBM-MSCs cultured on β-PVDF films coated with either collagen type I (a) or fibronectin (b) and corresponding morphological features. Nuclei are stained in blue, vinculin in green and F-actin in red. Results are expressed as mean ± SD (n=3). Only p-values <0.1 are shown. Scale bars represent 200 µm.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Fluorescence, Cell Culture, Staining

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: MYPT1 phosphorylation in hBM-MSCs cultured on collagen type I-coated (a) and fibronectin-coated (b) β-PVDF films of varying surface potential. Results are expressed as mean ± SD (n=3). Only p-values <0.1 are shown.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Phospho-proteomics, Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Fluorescence images of hBM-MSCs cultured on β-PVDF films coated with either collagen type I (a) or fibronectin (b) immunostained against YAP and counterstained with Hoechst 33342, and the corresponding quantifications (right panels). Scale bars represent 200 µm. Results are expressed as mean ± SD (n=3). Only p-values <0.1 are shown.

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Fluorescence, Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Volcano plots with the −log 10 (p-value) plotted against their respective log 2 (fold change) of genes differentially expressed in hBM-MSCs, Venn diagrams and histogram plots showing genes that are down- or upregulated (p<0.05) in hBM-MSCs cultured on the indicated surfaces for 24 hours (a) or four days (b). Circle area in a and b is proportional to the number of genes. GSEA of the cells cultured on the different surfaces for 24 hours (c) and 4 days (d).

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Cell Culture

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: (a) Immunostaining of hBM-MSCs cultured on the different β-PVDF surfaces with glutaraldehyde-crosslinked collagen type I coating and corresponding morphologic analysis (b). MYTP1 phosphorylation (c) and YAP translocation (d and e) in hBM-MSCs cultured on the substrates. Only p-values <0.1 are shown. Scale bars represent 200 µm. Results are expressed as mean ± SD (n=3).

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Immunostaining, Cell Culture, Phospho-proteomics, Translocation Assay

Journal: bioRxiv

Article Title: Surface potential as a strong early osteogenic trigger via mechanotransduction and calcium accumulation

doi: 10.64898/2026.04.26.720950

Figure Lengend Snippet: Immunofluorescent images (a) and corresponding morphologic analysis (b) of hBM-MSCs treated with the ROCK inhibitor Y-27632 for 24 hours. YAP immunolocalization (c) and translocation (d) in treated cells. Only p-values <0.1 are shown. Scale bars represent 300 µm. Results are expressed as mean ± SD (n=4).

Article Snippet: hBM-MSCs (Cytion, 300665) were seeded at an approximate density of 5.000 cells/cm 2 on the different surfaces in basal medium (DMEM; 11995-065, Gibco), 10% fetal bovine serum (FBS; 11560636, Gibco), 1% Glutamax (13462629, Gibco) and 1% antibiotics (11548876, Gibco), and after 24 hours the culture medium was replaced with osteogenic medium (Basal medium supplemented with 0,28 mM ascorbic acid (A4544, Sigma-Aldrich), 10 mM β-glycerol-2-phosphate (G9422, Sigma-Aldrich), and 10 nM dexamethasone (D4902, Sigma-Aldrich)).

Techniques: Translocation Assay

Journal: Cells

Article Title: Enrichment of Human Dermal Stem Cells from Primary Cell Cultures through the Elimination of Fibroblasts.

doi: 10.3390/cells12060949

Figure Lengend Snippet: Figure 1. Frequency of stem cells in DSC-fibroblast co-cultures. Dermal cells were cultured in stem cell medium for up to two weeks to form spheres. The spheres were enzymatically dissociated with Accutase™, and the percentage of DSCs (NGFRp75-positive cells) in the culture was determined via flow cytometry. (A) Boxplot displaying the median with 25th/75th percentiles. The cross (×) inside the box indicates the mean DSC frequency. Dots represent outliers. (B) The histogram shows the abundance distribution of DSC frequencies of individual donor cell strains divided into ranges of 5%. Results from 66 donors.

Article Snippet: For MACS® positive selection (labeling of DSCs), Neural Crest Stem Cell (NCSC) MicroBeads that are conjugated to anti-NGFRp75 antibodies (cat. #130-097-127; Miltenyi Biotec) were used according to the manufacturer’s protocol, with slight modifi- Cells 2023, 12, 949 6 of 20 cations: up to 1 × 107 cells were resuspended in buffer to a total volume of 160 μL (instead of 80 μL/1 × 107 cells); 40 μL of NCSC MicroBeads/1 × 107 cells (instead of 20 μL/1 × 107 cells) were added and incubated for 15 min at 4 ◦C in the dark; wash step with centrifugation at 300× g for 5 min; cell pellet was resuspended in 1 mL of buffer (instead of 500 μL), and sample was placed into the tube rack in the autoMACS® Pro Separator; the double-column positive separation program Posseld2 and the wash program Rinse were selected.

Techniques: Cell Culture, Cytometry

Journal: Cells

Article Title: Enrichment of Human Dermal Stem Cells from Primary Cell Cultures through the Elimination of Fibroblasts.

doi: 10.3390/cells12060949

Figure Lengend Snippet: Figure 3. Selective detachment of DSC-fibroblast co-cultures with 10–20% stem cells. Dermal cells were incubated with Accutase™for 1, 2, or 3 min or with trypsin-EDTA for 0.5, 1, or 2 min, and detached cells were collected. (A) Frequency of DSCs in samples measured using NGFRp75 staining. (B) Viability of total cells examined with propidium iodide. (C) Plot of recovery rate (x-axis) versus purity (y-axis) of detached cells at the individual incubation times. Purity: frequency of DSCs. Recovery rate: ratio of the absolute number of DSCs in the detached sample to the absolute number of DSCs before detachment. Dotted lines indicate the DSC frequency of control cells detached with Accutase™(light gray) or trypsin-EDTA (dark gray). Results from one representative donor cell strain.

Article Snippet: For MACS® positive selection (labeling of DSCs), Neural Crest Stem Cell (NCSC) MicroBeads that are conjugated to anti-NGFRp75 antibodies (cat. #130-097-127; Miltenyi Biotec) were used according to the manufacturer’s protocol, with slight modifi- Cells 2023, 12, 949 6 of 20 cations: up to 1 × 107 cells were resuspended in buffer to a total volume of 160 μL (instead of 80 μL/1 × 107 cells); 40 μL of NCSC MicroBeads/1 × 107 cells (instead of 20 μL/1 × 107 cells) were added and incubated for 15 min at 4 ◦C in the dark; wash step with centrifugation at 300× g for 5 min; cell pellet was resuspended in 1 mL of buffer (instead of 500 μL), and sample was placed into the tube rack in the autoMACS® Pro Separator; the double-column positive separation program Posseld2 and the wash program Rinse were selected.

Techniques: Incubation, Staining, Control

Journal: Cells

Article Title: Enrichment of Human Dermal Stem Cells from Primary Cell Cultures through the Elimination of Fibroblasts.

doi: 10.3390/cells12060949

Figure Lengend Snippet: Figure 5. Positive selection (labeling of DSCs). (A–C) EasySep™column-free positive selection with the EasySep™Human CD271 Positive Selection Kit II (StemCell Technologies). Sample size: n = 6. (D–F) MACS® automatic column-based positive selection with Neural Crest Stem Cell (NCSC) MicroBeads and the autoMACS® Pro Separator (Miltenyi Biotec). Program: Posseld2. Sample size: n = 13. Values are presented as mean ± SDs. Pre: initial sample, neg frac: fibroblast fraction, pos frac: DSC fraction, d11–12: 11–12 days of cultivation. Frequency of DSCs in the separate fractions was determined via flow cytometry analysis of NGFRp75 (y-axis of dot plots) and CD90 (x-axis of dot plots). Following separation, the enriched DSC fraction was cultivated in stem cell medium. After 11–12 days, the proportion of DSCs in culture was measured again via flow cytometry, and cells were additionally stained immunohistochemically for NGFRp75 (red) and CD90 (green). Nuclei were counterstained with DAPI (blue). Scale bars: 200 µm.

Article Snippet: For MACS® positive selection (labeling of DSCs), Neural Crest Stem Cell (NCSC) MicroBeads that are conjugated to anti-NGFRp75 antibodies (cat. #130-097-127; Miltenyi Biotec) were used according to the manufacturer’s protocol, with slight modifi- Cells 2023, 12, 949 6 of 20 cations: up to 1 × 107 cells were resuspended in buffer to a total volume of 160 μL (instead of 80 μL/1 × 107 cells); 40 μL of NCSC MicroBeads/1 × 107 cells (instead of 20 μL/1 × 107 cells) were added and incubated for 15 min at 4 ◦C in the dark; wash step with centrifugation at 300× g for 5 min; cell pellet was resuspended in 1 mL of buffer (instead of 500 μL), and sample was placed into the tube rack in the autoMACS® Pro Separator; the double-column positive separation program Posseld2 and the wash program Rinse were selected.

Techniques: Selection, Labeling, Cytometry, Staining

Journal: Cells

Article Title: Enrichment of Human Dermal Stem Cells from Primary Cell Cultures through the Elimination of Fibroblasts.

doi: 10.3390/cells12060949

Figure Lengend Snippet: Figure 6. Overview of purity and recovery of DSCs after the individual selection methods. (A) Purity. Frequency of DSCs in the purified fraction after separation determined via flow cytometry analysis of NGFRp75. Values are presented as mean ± SDs. (B) Recovery rate. Ratio of the absolute number of DSCs in the purified fraction after separation to the absolute number of DSCs in the pre sample. Values are presented as mean ± SDs. (C) Plot of recovery rate (x-axis) versus purity (y-axis) of the individual selection methods. Sample size: a. n = 5; b. n = 3; c. n = 2; d. n = 3; e. n = 3; f. n = 2; g. n = 6; h. n = 13.

Article Snippet: For MACS® positive selection (labeling of DSCs), Neural Crest Stem Cell (NCSC) MicroBeads that are conjugated to anti-NGFRp75 antibodies (cat. #130-097-127; Miltenyi Biotec) were used according to the manufacturer’s protocol, with slight modifi- Cells 2023, 12, 949 6 of 20 cations: up to 1 × 107 cells were resuspended in buffer to a total volume of 160 μL (instead of 80 μL/1 × 107 cells); 40 μL of NCSC MicroBeads/1 × 107 cells (instead of 20 μL/1 × 107 cells) were added and incubated for 15 min at 4 ◦C in the dark; wash step with centrifugation at 300× g for 5 min; cell pellet was resuspended in 1 mL of buffer (instead of 500 μL), and sample was placed into the tube rack in the autoMACS® Pro Separator; the double-column positive separation program Posseld2 and the wash program Rinse were selected.

Techniques: Selection, Cytometry

Journal: Cell Death & Disease

Article Title: Zebularine regulates early stages of mESC differentiation: effect on cardiac commitment

doi: 10.1038/cddis.2013.88

Figure Lengend Snippet: Characterization of HS-181 cell line after treatment with zebularine. ( a ) RT-PCR of cardiac markers after treatment with zebularine. Myh7, Myh6, Actc, cTnI and Serca2 present more expression after treatment. ( b ) Immunostaining of treated cells to detect cardiac-specific proteins. Scale bars; 50 μ m. ( c ) Blots of Proteome Profiler Array and the resulting quantification histograms demonstrating inhibition of pluripotency marker expression and ( d ) increased levels of mesodermic proteins after zebularine treatment (black arrows)

Article Snippet: Protein expression profiles were assayed using the specific human pluripotent Stem Cell array kit ‘Proteome Profiler Array' (R&D Systems Europe, Abingdon, UK; ARY010) following the manufacturer's instructions.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Immunostaining, Inhibition, Marker

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: Expression of neural and neurotrophic markers in nucleus pulposus cells isolated from degenerated intervertebral disc.

doi: 10.1002/jor.22098

Figure Lengend Snippet: Figure 2. Mesenchymal Profile of Human NPs-IVD: (A) Immunophenotypic profile, by citofluorimetric analysis, of nucleus pulposus from 14 human degenerated intervertebral discs. (B) NPs-IVD show mesenchymal properties under chondrogenic, adipogenic, and osteogenic differentiation.

Article Snippet: Human Mesenchymal Stem Cell Functional Identification Kit (R&D Systems, Minneapolis, MN) was used to induce adipogenic, chondrogenic, and osteogenic differentiation.

Techniques:

Journal: Molecular Therapy. Nucleic Acids

Article Title: Ratio of miRNA-29 to miRNA-199 expression coordinates mesenchymal stem cell repair of bleomycin-induced pulmonary injury

doi: 10.1016/j.omtn.2025.102461

Figure Lengend Snippet: Bleomycin-induced lung injury, which is reduced after adipose-derived mesenchymal stem cell instillation (A) Representative μCT transverse and coronal lung sections acquired from aged (22-month-old) male C57BL/6 mice at baseline (left) and 7 days following intratracheal bleomycin (BLM, 2.0 U/kg) administration (right) demonstrating increased lung density and loss of airspaces. (B) Saline treatment did not result in evidence of lung injury on μCT scan at baseline (left) or 7 days post-instillation (right). Histological sections of lung tissue collected at day 21 post-BLM were stained with Masson’s trichrome as described in . (C and D) Representative photomicrographs (20× and 40× magnifications) of lung sections from saline-treated control mice (C) and BLM-treated mice (D). (E) Infusion of adipose-derived mesenchymal stem cells (ASCs) 12 days post-BLM instillation resulted in reduced severity of pulmonary fibrosis (PF). (F) Degree of PF on histological sections was measured by semi-quantitative Ashcroft score as described in . BLM-induced lung injury resulted in increased Ashcroft score compared to saline controls. Infusion with ASCs 12 days post-BLM injury resulted in decreased Ashcroft score. (G) Intratracheal BLM instillation increased lung collagen content as measured by hydroxyproline assays as described in . Mice treated with ASCs on day 12 post-BLM had decreased lung collagen content compared to BLM-only controls. Each data point represents an individual biological replicate (mouse); n = 6–10 mice/group. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. (H) Ratio of pAKT to AKT protein expression in lung tissue of mice was quantified by western blot analysis at day 21 post-BLM sacrifice. Aged C57BL/6 mice treated with intratracheal BLM demonstrated increased pAKT/AKT protein expression compared to saline-treated controls. Lungs from mice treated with intravenous infusion of ASCs 12 days post-BLM-induced injury demonstrated decreased expression of pAKT/AKT compared to BLM-only group. Inset shows a representative western blot and β-actin loading control. Data are graphed as individual biological replicates ( n = 6–8 mice/group); ∗ p < 0.05.

Article Snippet: For mesenchymal differentiation potential, the Mouse Mesenchymal Stem Cell Functional Identification Kit (R&D Systems, Minneapolis, MN) was used according to the manufacturer’s instructions.

Techniques: Derivative Assay, Saline, Staining, Control, Expressing, Western Blot

Journal: Stem cell research

Article Title: Generation of two familial hypercholesterolemia patient-specific induced pluripotent stem cell lines harboring heterozygous mutations in the LDLR gene

doi: 10.1016/j.scr.2024.103463

Figure Lengend Snippet:

Article Snippet: Ectoderm was inducted with the Human Pluripotent Stem Cell Functional Identification Kit (R&D Systems #SC027B).

Techniques: Virus, Modification

Journal: STAR Protocols

Article Title: An optimized chromatin immunoprecipitation protocol using Staph-seq for analyzing genome-wide protein-DNA interactions

doi: 10.1016/j.xpro.2022.101918

Figure Lengend Snippet:

Article Snippet: V6.5 mouse embryonic stem cells , Novus Biologicals , Cat#NBP1-41162.

Techniques: Recombinant, SYBR Green Assay, Next-Generation Sequencing, Agarose Gel Electrophoresis, Purification, Software, Real-time Polymerase Chain Reaction