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: 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: Heliyon

Article Title: MSCs-laden silk Fibroin/GelMA hydrogels with incorporation of platelet-rich plasma for chondrogenic construct

doi: 10.1016/j.heliyon.2023.e14349

Figure Lengend Snippet: A schematic illustration of the preparation of MSCs-laden gelatin methacrylate/silk fibroin (GelMA/SF, SG) hydrogel with the incorporation of PRP for treating KOA to reconstruct cartilage.

Article Snippet: Due to the potential differentiation into cartilage cells (chondrocytes), bone marrow-derived mesenchymal stem cells (MSCs, Normal, Human, ATCC®PCS-500-012TM) were used in this work.

Techniques:

Journal: Heliyon

Article Title: MSCs-laden silk Fibroin/GelMA hydrogels with incorporation of platelet-rich plasma for chondrogenic construct

doi: 10.1016/j.heliyon.2023.e14349

Figure Lengend Snippet: The MSCs proliferation after loaded in SG hydrogels and cultured for 72 h, cells cultured without hydrogel as a control group. Laser scanning confocal microscope (LSCM) images of MSCs loaded in SG-1 ( A , B , and C ), SG-2 (D, E, and F), and SG-3 ( G , H , and I ) hydrogels, and without hydrogels ( J , K , and L ) cultured for 72 h, images ( A , D , G , and J ) displayed that cell nucleus stained using DAPI as shown in blue color, images ( B , E , H , and K ) displayed that cytoskeleton stained using Phalloidin-FITC as shown in green color, and image ( C , F , I , and L ) was merged from DAPI, and Phalloidin-FITC stained images, respectively. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Due to the potential differentiation into cartilage cells (chondrocytes), bone marrow-derived mesenchymal stem cells (MSCs, Normal, Human, ATCC®PCS-500-012TM) were used in this work.

Techniques: Cell Culture, Control, Microscopy, Staining

Journal: Heliyon

Article Title: MSCs-laden silk Fibroin/GelMA hydrogels with incorporation of platelet-rich plasma for chondrogenic construct

doi: 10.1016/j.heliyon.2023.e14349

Figure Lengend Snippet: Cell viability of MSCs after loaded in SG hydrogels and cultured for different days. * p < 0.05, and ** p < 0.01.

Article Snippet: Due to the potential differentiation into cartilage cells (chondrocytes), bone marrow-derived mesenchymal stem cells (MSCs, Normal, Human, ATCC®PCS-500-012TM) were used in this work.

Techniques: Cell Culture

Journal: Heliyon

Article Title: MSCs-laden silk Fibroin/GelMA hydrogels with incorporation of platelet-rich plasma for chondrogenic construct

doi: 10.1016/j.heliyon.2023.e14349

Figure Lengend Snippet: Optical micrographs of ( A-H ) H&E and ( I–P ) Masson staining slices of tissues after SG hydrogels ( C, D, K, and L ), SG hydrogels incorporated with PRP ( E, F, M, and N ), and MSCs-laden SG hydrogels incorporated with PRP ( G, H, O, and P ) were intra-articular injected into SD rats for 8 weeks, respectively, the SD rats without hydrogels intervention as the control group ( A, B, I, and J ).

Article Snippet: Due to the potential differentiation into cartilage cells (chondrocytes), bone marrow-derived mesenchymal stem cells (MSCs, Normal, Human, ATCC®PCS-500-012TM) were used in this work.

Techniques: Staining, Injection, Control

Journal: Heliyon

Article Title: MSCs-laden silk Fibroin/GelMA hydrogels with incorporation of platelet-rich plasma for chondrogenic construct

doi: 10.1016/j.heliyon.2023.e14349

Figure Lengend Snippet: The obtained scores for SG hydrogel, SG hydrogel with PRP, MSCs-loaded hydrogel with PRP, and control group using Mankin Scoring system. * p < 0.05, and ** p < 0.01.

Article Snippet: Due to the potential differentiation into cartilage cells (chondrocytes), bone marrow-derived mesenchymal stem cells (MSCs, Normal, Human, ATCC®PCS-500-012TM) were used in this work.

Techniques: Control

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 1. MSCs synergized with A1AT to modulate inflammation in Raw 264.7 macrophages. Cells were stimulated with 100 ng/mL LPS plus 10 ng/mL IFNγ and treated with 0.5 mg/mL A1AT or MSCs (MSC/MΦ = 1/10) or their combination. Dexamethasone (Dex, 1 µg/mL) was used as a benchmark. Pro-inflammatory mouse cytokine IL6 (A), TNFα (B), and anti-inflammatory mouse cytokine IL10 (C) were measured via ELISA. The IL6/IL10 (D) and TNFα/IL10 ratio (E) was also shown. *:p < 0.05, **:p < 0.01, ***:p < 0.001.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 2. MSCs synergized with A1AT to modulate inflammation in primary human PBMCs. Cells were stimulated with 100 ng/mL LPS + anti-CD3/CD28 antibodies (positive) and treated with 0.5 mg/mL A1AT or MSCs (MSC/PBMC = 1/10) or their combination for 24 hs. Dexamethasone (Dex, 1 µg/mL) was used as a benchmark. PBMCs without activation and treatment were used as a negative control. Pro-inflammatory human cytokine IL6 (A), TNFα (B), and anti-inflammatory human cytokine IL10 (C) were measured via ELISA. The IL6/IL10 (D) and TNFα/IL10 ratio (E) was also shown. *:p < 0.05, **:p < 0.01, ***:p < 0.001.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: Activation Assay, Negative Control, Enzyme-linked Immunosorbent Assay

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 3. MSCs and A1AT combination treatment reduced neutrophil ROS and NETs production. HL-60 cells derived neutrophils were stimulated with 100 nM PMA and treated with 0.5 mg/mL A1AT or MSCs (MSC/neutrophil = 1/10) or their combination for 4 hs. Reactive oxygen species (ROS) (A-B) and neutrophil extracellular traps (NETs) production (C-D) were analyzed. *:p < 0.05, **:p < 0.01, ***:p < 0.001.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: Derivative Assay

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 4. MSC and A1AT combination treatment enhanced phagocytosis in THP-1 derived macrophages (A-D) and HL-60 cells derived neutrophils (E-H). Macrophages were stimulated with 100 ng/mL LPS plus 10 ng/mL IFNγ for 24 hs. Neutrophils were stimulated with 100 nM PMA for 4 hs. Cells were treated with 0.5 mg/mL A1AT or MSCs

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: Derivative Assay

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 5. MSCs synergized with A1AT to improve survival rate and reduce lung injury in mice. (A) Illustration of the model. (B) The survival rate and (C) body weight development. N = 6. (D) H&E staining and (E) lung injury scores. The lung injury scores were calculated based on the five criteria shown in (D). *:p < 0.05, **:p < 0.01, ***:p < 0.001.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: Staining

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 6. MSCs and A1AT synergized in reducing total protein (A) and pro-inflammatory cytokines while increasing anti-inflammatory cytokine IL10 (B-F) in BALF. *:p < 0.05, **:p < 0.01, ***:p < 0.001.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques:

Journal: Theranostics

Article Title: Mesenchymal stromal cells and alpha-1 antitrypsin have a strong synergy in modulating inflammation and its resolution.

doi: 10.7150/thno.83942

Figure Lengend Snippet: Figure 8. MSCs synergized with A1AT to reduce total cell (A-B), macrophage (C), and neutrophil number (D) in BALF. The M1/M2 macrophage ratio was reduced by all treatments (E). *:p < 0.05, **:p < 0.01, ***:p < 0.001. (F-G) MSCs synergized with A1AT to reduce cell death as identified via TUNEL staining.

Article Snippet: P4 MSCs were characterized with the Human Mesenchymal Stem Cell Verification Flow Kit (R&D Systems), including antibodies for positive markers CD90, CD73, CD105, and negative markers CD45, CD34, CD11b, CD79A, HLA-DR, as well as the Human Mesenchymal Stem Cells Multi-Color Flow Kit (R&D Systems) including antibodies for positive markers CD44, CD106, CD146, and CD166.

Techniques: TUNEL Assay, Staining

Journal: Cells

Article Title: hTERT-Immortalized Mesenchymal Stem Cell-Derived Extracellular Vesicles: Large-Scale Manufacturing, Cargo Profiling, and Functional Effects in Retinal Epithelial Cells

doi: 10.3390/cells13100861

Figure Lengend Snippet: Nanoparticle tracking analysis and EV morphology. NTA was performed on two biological replicates (i.e., Lot 1 and Lot 2) of ( a ) hTERT-immortalized MSC EVs and ( b ) PC3 EVs to analyze relative concentration (particles/mL) and size distribution. Dashed lines indicate values of 50 nm and 200 nm. ( c ) The average % of EVs from each lot that measured between 50–200 nm in size and 200+ nm in size. n = 6 (2 biological replicates and each biological replicate was measured in triplicate). ** = p < 0.01. Representative TEM images show the appearance of ( d ) hTERT-immortalized MSC EVs and ( e ) PC 3 EVs. Scale bar = 100 nm.

Article Snippet: The EVs used in this study are available from ATCC as follows: hTERT -immortalized MSC (ATCC ® SCRC-4000-EXM™), PC3 (ATCC ® CRL-1435-EXM™), and HCT 116 (ATCC ® CCL-247™).

Techniques: Concentration Assay

Journal: Cells

Article Title: hTERT-Immortalized Mesenchymal Stem Cell-Derived Extracellular Vesicles: Large-Scale Manufacturing, Cargo Profiling, and Functional Effects in Retinal Epithelial Cells

doi: 10.3390/cells13100861

Figure Lengend Snippet: Characterization of EVs. ( a ) Western blot for CD63, CD9, CD81, and GAPDH from two independent lots (biological replicates) of hTERT-immortalized MSC EVs. ( b ) Western blot for CD63, CD9, CD81, and GAPDH from two independent lots (biological replicates) of PC3 EVs. Multiplex analysis was performed to analyze tetraspanins (CD63, CD81, and CD9). ( c ) Average ECL values from two independent lots (biological replicates) of hTERT-immortalized MSC EVs. Each lot was assayed in duplicate. ( d ) Average ECL values from two independent lots (biological replicates) of PC3 EVs. Each lot was assayed in duplicate. (* p < 0.05; ** p < 0.01; **** p < 0.0001).

Article Snippet: The EVs used in this study are available from ATCC as follows: hTERT -immortalized MSC (ATCC ® SCRC-4000-EXM™), PC3 (ATCC ® CRL-1435-EXM™), and HCT 116 (ATCC ® CCL-247™).

Techniques: Western Blot, Multiplex Assay

Journal: Cells

Article Title: hTERT-Immortalized Mesenchymal Stem Cell-Derived Extracellular Vesicles: Large-Scale Manufacturing, Cargo Profiling, and Functional Effects in Retinal Epithelial Cells

doi: 10.3390/cells13100861

Figure Lengend Snippet: EV surface marker profiling and cell migration Assay. Multiplex analysis was performed to analyze various EV-associated surface marker proteins. Assays were run in duplicate. The average ECL values of each surface marker were analyzed and compared between hTERT-immortalized MSC and PC3 EVs. ( a ) THY-1, Thrombomodulin, and Neprilysin were significantly enriched in hTERT-immortalized MSC EVs. ( b ) MCAM, EPCAM, ICAM-1, EGFR, and ALCAM were significantly enriched in PC3 EVs. **** p < 0.0001; *** p < 0.001; ** p < 0.01. ( c ) A cell migration assay was performed using RPE cells. After the creation of an artificial gap, cells were immediately treated with EVs at an approximate ratio of 1:10,000 (recipient cell:EV). Representative images display gap closure over 48 h in response EVs. Scale bar = 1000 µm. ( d ) GraphPad Prism was used to analyze raw images to quantify percent gap coverage. **** p < 0.0001 relative to untreated.

Article Snippet: The EVs used in this study are available from ATCC as follows: hTERT -immortalized MSC (ATCC ® SCRC-4000-EXM™), PC3 (ATCC ® CRL-1435-EXM™), and HCT 116 (ATCC ® CCL-247™).

Techniques: Marker, Cell Migration Assay, Multiplex Assay

Journal: Cells

Article Title: hTERT-Immortalized Mesenchymal Stem Cell-Derived Extracellular Vesicles: Large-Scale Manufacturing, Cargo Profiling, and Functional Effects in Retinal Epithelial Cells

doi: 10.3390/cells13100861

Figure Lengend Snippet: Profiling of EV-associated cargo. ( a ) The results from MS show the number of unique and shared proteins between hTERT-immortalized MSC EVs and PC3 EVs. ( b ) The STRING database was used to calculate the PPIs of peptides extracted from each EV preparation. Proteins are represented by nodes that are connected by lines representative of their confidence level. Colors represent specific functions that are associated with proteins. ( c ) Total RNA was isolated from hTERT-immortalized MSCs, and RT-qPCR was performed to target candidate mRNAs. Average Cq values are shown. n = 3. ( d ) Total RNA was isolated from hTERT-immortalized MSC EVs, and RT-qPCR was performed to target candidate mRNAs. Average Cq values are shown. n = 3. ( e ) EVs were assayed for the presence of inflammatory cytokines using the S-PLEX ® proinflammatory Panel 1 Kit (Meso Scale Diagnostics) following the manufacturer’s instructions. Assays were run in duplicate. Analyte concentrations were calculated using a 4-PL fit of a standard curve of the calibrators. The data shown represent samples assayed without lysis. *** p < 0.001; * p < 0.05.

Article Snippet: The EVs used in this study are available from ATCC as follows: hTERT -immortalized MSC (ATCC ® SCRC-4000-EXM™), PC3 (ATCC ® CRL-1435-EXM™), and HCT 116 (ATCC ® CCL-247™).

Techniques: Isolation, Quantitative RT-PCR, Lysis