Journal: Stem Cells International

Article Title: Interleukin-1 β Enhances Umbilical Cord Mesenchymal Stem Cell Adhesion Ability on Human Umbilical Vein Endothelial Cells via LFA-1/ICAM-1 Interaction

doi: 10.1155/2019/7267142

Figure Lengend Snippet: IL-1 β induces increased expression of LFA-1 in MSCs. (a) MSCs without treated IL-1 β as a control group and treated with IL-1 β for 15, 30, 120, and 360 minutes (scale bar: 50 μ m). Histograms analyze LFA-1 (green) fluorescence intensity and location of one single MSC. White blocks show a single MSC. Red lines show the histogram detection sites. Results of measurement of LFA-1 (green) and cell nucleus (blue) fluorescence intensity are quantified by MetaMorph. (b) Example of Western blot results of the LFA-1 (129 kDa) expression in cytosolic and membrane fractions. Cells were treated with IL-1 β for 15, 30, and 120 minutes. (c) Quantitative graphs of the Western blot results of LFA-1 protein expression of (b) ( n = 3, ∗∗ P < 0.01). (d) Example of Western blot results of the LFA-1 (129 kDa) from cell membrane protein. Cells treated with IL-1 β and IL-1RA or cotreated with IL-1 β and IL-1RA. (e) Quantitative graphs of the Western blot results of LFA-1 protein expression of (d) ( n = 3, ∗∗∗ P < 0.005, and ∗∗ P < 0.01).

Article Snippet: Umbilical cord mesenchymal stem cells (UC-MSCs) were purchased from the Bioresource Collection and Research Center, Hsinchu, Taiwan.

Techniques: Expressing, Control, Fluorescence, Western Blot, Membrane

Journal: Stem Cells International

Article Title: Interleukin-1 β Enhances Umbilical Cord Mesenchymal Stem Cell Adhesion Ability on Human Umbilical Vein Endothelial Cells via LFA-1/ICAM-1 Interaction

doi: 10.1155/2019/7267142

Figure Lengend Snippet: IL-1 β -induced ICAM-1 expression in HUVECs and enhanced MSC adhesion to HUVECs via LFA-1/ICAM-1. (a) Immunocytochemistry staining of ICAM-1 (green) and cell nucleus (blue) in HUVECs treated with or without IL-1 β for 15, 30, 120, and 360 minutes (scale bar: 50 μ m). Histograms analyze ICAM-1 (green) fluorescence intensity and location of one single MSC. White blocks show a single MSC. Red lines show histogram detection sites. Results of measured ICAM-1 (green) and cell nucleus (blue) fluorescence intensity are quantified by MetaMorph. (b) Representative image of the adhesion of MSCs to HUVECs. MSCs were treated with IL-1 β , IL-1RA, and adhesion to IL-1 β -activated HUVECs or nonactivated HUVECs. MSCs labelled with calcein AM (5 μ M) (green), HUVECs, and MSC cell nuclei were stained with Hoechst 33258 (blue) (scale bar: 50 μ m). (c) Quantitative graphs of the cell adhesion assay results of MSC adhesion to nonactivated HUVECs (black bars) or IL-1 β -activated HUVECs (gray bars). Values were the cell number fold change relative to the control group. Data represent mean ± SD ( ∗∗∗ P < 0.005, ∗∗ P < 0.01, and ∗ P < 0.05) (N.S.: nonsignificance). (d) Representative image of MSC adhesion on HUVECs. Cell adhesion assay to determine the percentage of MSC adhesion on the HUVEC monolayer. MSC adhesion to IL-1 β -activated HUVECs or nonactivated HUVECs after treatment with IL-1 β and the inhibitor lovastatin. (e) Quantitative graphs of the cell adhesion assay results of MSC adhesion to nonactivated HUVECs (black bars) or IL-1 β -activated HUVECs (gray bars). MSCs treated with IL-1 β and the inhibitor lovastatin. Data represent mean ± SD ( n = 3, ∗∗∗ P < 0.005, ∗∗ P < 0.01, and ∗ P < 0.05).

Article Snippet: Umbilical cord mesenchymal stem cells (UC-MSCs) were purchased from the Bioresource Collection and Research Center, Hsinchu, Taiwan.

Techniques: Expressing, Immunocytochemistry, Staining, Fluorescence, Cell Adhesion Assay, Control

Journal: Stem Cells International

Article Title: Interleukin-1 β Enhances Umbilical Cord Mesenchymal Stem Cell Adhesion Ability on Human Umbilical Vein Endothelial Cells via LFA-1/ICAM-1 Interaction

doi: 10.1155/2019/7267142

Figure Lengend Snippet: Effects of cycloheximide in MSC adhesion to HUVECs. (a) Western blot results of the LFA-1 (129 kDa) from membrane protein. Cells treated with IL-1 β and cycloheximide (CYH) or cotreated with both IL-1 β and CYH. (b) Quantitative graphs of the Western blot results of LFA-1 protein expression of (a). ( n = 3, ∗∗∗ P < 0.01, and ∗ P < 0.05). (c) Representative image of MSC adhesion on HUVECs. MSCs treated with IL-1 β ; cycloheximide adhesion to IL-1 β -activated HUVECs or nonactivated HUVECs. MSCs labelled with calcein AM (5 μ M) (green), HUVECs, and MSC cell nuclei were stained with Hoechst 33258 (blue) (scale bar: 50 μ m). (d) Quantitative graphs of the cell adhesion assay results of MSC adhesion to nonactivated HUVECs (black bars) or IL-1 β -activated HUVECs (gray bars). MSCs treated with IL-1 β and cycloheximide. MSC adhesion cell number counted in five randomly selected fields in a single representative experiment performed three times. Values were the cell number fold change relative to the control group. Data represent mean ± SD ( n = 3, ∗∗∗ P < 0.005, and ∗∗ P < 0.01) (N.S.: nonsignificance).

Article Snippet: Umbilical cord mesenchymal stem cells (UC-MSCs) were purchased from the Bioresource Collection and Research Center, Hsinchu, Taiwan.

Techniques: Western Blot, Membrane, Expressing, Staining, Cell Adhesion Assay, Control

Journal: Stem Cells International

Article Title: Interleukin-1 β Enhances Umbilical Cord Mesenchymal Stem Cell Adhesion Ability on Human Umbilical Vein Endothelial Cells via LFA-1/ICAM-1 Interaction

doi: 10.1155/2019/7267142

Figure Lengend Snippet: p38 MAPK signaling pathway is involved in IL-1 β -mediated LFA-1 expression in MSCs and MSC adhesion to HUVECs. (a) Immunocytochemistry staining for LFA-1 (green) and DAPI (blue) in MSCs. Cells were treated with p38 MAPK (SB 203580, 5 μ M), AKT (GSK690693, 20 μ M), ERK1/2 (U0126 20 μ M), and JNK (SP600125 20 nM) inhibitor and combined with IL-1 β for 30 minutes (scale bar: 50 μ m). (b) Western blot results of the p38 MAPK and phosphorylated p38 MAPK from the lysates of cells pretreated with IL-1 β at 0, 1, 3, 5, 10, and 20 minutes. (c) Western blot results of the LFA-1 (129 kDa) expression in membrane fractions of MSCs treated with IL-1 β and SB 203580 (SB) or cotreated with both IL-1 β and SB. (d) Quantitative graphs of the Western blot results of LFA-1 expression of (c) ( n = 3, ∗∗ P < 0.01). (e) Representative image of MSC adhesion on HUVECs. MSCs were treated with IL-1 β and inhibitor SB 203580 adhesion to IL-1 β -activated HUVECs or nonactivated HUVECs. MSCs labelled with calcein AM (5 μ M) (green), HUVECs, and MSC cell nuclei were stained with Hoechst 33258 (blue) (scale bar: 50 μ m). (f) Quantitative graphs of the cell adhesion assay results of MSCs treated with IL-1 β and MAPK inhibitor SB 203580 adhesion to nonactivated HUVECs (black bars) or IL-1 β -activated HUVECs (gray bars). Values were the cell number fold change relative to the control group. Data represent mean ± SD ( n = 3, ∗∗∗ P < 0.005, ∗∗ P < 0.01, and ∗ P < 0.05) (N.S.: nonsignificance).

Article Snippet: Umbilical cord mesenchymal stem cells (UC-MSCs) were purchased from the Bioresource Collection and Research Center, Hsinchu, Taiwan.

Techniques: Expressing, Immunocytochemistry, Staining, Western Blot, Membrane, Cell Adhesion Assay, Control

Journal: Stem Cells International

Article Title: Interleukin-1 β Enhances Umbilical Cord Mesenchymal Stem Cell Adhesion Ability on Human Umbilical Vein Endothelial Cells via LFA-1/ICAM-1 Interaction

doi: 10.1155/2019/7267142

Figure Lengend Snippet: Schematic diagram of IL-1 β signaling pathway in MSC adhesion to HUVECs. The schematic diagram depicts the proposed role of IL-1 β signaling pathway in MSC adhesion to HUVECs. The process of cell adhesion is initiated by IL-1 β through p38 MAPK; induced expression of LFA-1 in MSCs enhances the cell adhesion to IL-1 β -induced ICAM-1 in HUVECs.

Article Snippet: Umbilical cord mesenchymal stem cells (UC-MSCs) were purchased from the Bioresource Collection and Research Center, Hsinchu, Taiwan.

Techniques: Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: All-trans retinoic acid pretreatment of mesenchymal stem cells enhances the therapeutic effect on acute kidney injury

doi: 10.1186/s12964-024-01671-1

Figure Lengend Snippet: ATRA pretreatment improves the protective effect of MSCs on renal function and pathologic damage in I/R-induced AKI. a GFR assessment using transcutaneous clearance of FITC-sinistrin ( n = 5). b, c SCr and BUN levels ( n = 6). d Semi-quantitative analysis of histological features ( n = 6). e Representative images of PAS staining on renal tissue sections in different groups of mice. Scale bars, 200 μm and 50 μm in scanned and zoom in figures, respectively. f Representative immunofluorescence staining of KIM-1. Scale bars, 200 μm and 50 μm in scanned and zoom in figures, respectively. g, h Representative western blot images and quantitative analysis of KIM-1 expression in kidney tissues ( n = 3). Data are presented as the means ± SD. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05

Article Snippet: Human umbilical cord mesenchymal stem cells (hUC-MSCs) were acquired from the Beijing Zhongyuan Company (Beijing, China).

Techniques: Staining, Immunofluorescence, Western Blot, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: All-trans retinoic acid pretreatment of mesenchymal stem cells enhances the therapeutic effect on acute kidney injury

doi: 10.1186/s12964-024-01671-1

Figure Lengend Snippet: ATRA regulates the expression of HA by regulating the binding of the transcription factors RARα/RXRγ to HAS2 gene. a Viability of MSCs after ATRA pretreatment ( n = 6). b Volcano plot of showing the differentially expressed genes in ATRA-MSCs versus DMSO-MSCs ( n = 3). Red dots indicate significant up-regulation, green dots indicate significant down-regulation, and gray dots indicate insignificant differences. c Scatter plot of GO enrichment analysis ( n = 3). The number of enriched differential genes can be observed by the dot size, and the significance of the enrichment is indicated by the dot color intensity. d The mRNA expression of HAS1, HAS2 and HAS3 ( n = 3). e Transfection efficiency of different siRNA ( n = 5). f The concentration of HA in the MSCs culture supernatant ( n = 4). g RARα/RXRγ binding motif logo generated by the WebLogo tool. h Schematic representation of possible binding sites for the transcription factor RARα/RXRγ in the HAS2 promoter region. i ChIP-qPCR analysis of HAS2 target promotor regions ( n = 3). Data are presented as the means ± SD. **** P < 0.0001, ** P < 0.01

Article Snippet: Human umbilical cord mesenchymal stem cells (hUC-MSCs) were acquired from the Beijing Zhongyuan Company (Beijing, China).

Techniques: Expressing, Binding Assay, Transfection, Concentration Assay, Generated, ChIP-qPCR

Journal: Cell Communication and Signaling : CCS

Article Title: All-trans retinoic acid pretreatment of mesenchymal stem cells enhances the therapeutic effect on acute kidney injury

doi: 10.1186/s12964-024-01671-1

Figure Lengend Snippet: ATRA enhances the anti-inflammatory, anti-apoptosis and pro-proliferation effects of MSCs on H/R-induced HK-2 cells in vitro. a-g Representative western blot images and quantitative analysis of TNF-a, IL-6, Bax, Bcl-2, and cleaved caspase-3 in HK-2 cells ( n = 3). h, I Representative western blot images and quantitative analysis of PCNA in HK-2 cells ( n = 3). Data are presented as the means ± SD. *** P < 0.001, ** P < 0.01, * P < 0.05

Article Snippet: Human umbilical cord mesenchymal stem cells (hUC-MSCs) were acquired from the Beijing Zhongyuan Company (Beijing, China).

Techniques: In Vitro, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: All-trans retinoic acid pretreatment of mesenchymal stem cells enhances the therapeutic effect on acute kidney injury

doi: 10.1186/s12964-024-01671-1

Figure Lengend Snippet: ATRA enhances the anti-inflammatory, anti-apoptosis and pro-proliferation effects of MSCs on I/R-induced AKI in vivo. a TNF-a mRNA expression level ( n = 3). b IL-6 mRNA expression level ( n = 3). c IL-1β mRNA expression level ( n = 3). d-j Representative western blot images and quantitative analysis of TNF-a, IL-6, Bax, Bcl-2, cleaved caspase-3 and PCNA in kidney tissues ( n = 3). k Apoptotic cells were labeled with the TUNEL assay kit (red) with nuclei counterstained with DAPI (blue) in kidney sections. Scale bar, 50 μm. l Quantitative analysis of the percentage of apoptotic cells ( n = 3). m Representative images of PCNA in kidney tissues. Scale bar, 40 μm. n Quantitative analysis of PCNA-positive cells ( n = 3). Data are presented as the means ± SD. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05

Article Snippet: Human umbilical cord mesenchymal stem cells (hUC-MSCs) were acquired from the Beijing Zhongyuan Company (Beijing, China).

Techniques: In Vivo, Expressing, Western Blot, Labeling, TUNEL Assay

Journal: Cell Communication and Signaling : CCS

Article Title: All-trans retinoic acid pretreatment of mesenchymal stem cells enhances the therapeutic effect on acute kidney injury

doi: 10.1186/s12964-024-01671-1

Figure Lengend Snippet: Inhibition of the HA/CD44 axis reverses the anti-inflammatory, anti-apoptotic and pro-proliferative effects of ATRA-MSCs. a, c-i Western blot images and quantitative densitometry analysis of TNF-a, IL-6, Bax, Bcl-2 and cleaved caspase-3 in HK-2 cells ( n = 3). b, j-l Western blot images and quantitative densitometry analysis of p-AKT and total AKT in HK-2 cells ( n = 3). Data are presented as the means ± SD. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05

Article Snippet: Human umbilical cord mesenchymal stem cells (hUC-MSCs) were acquired from the Beijing Zhongyuan Company (Beijing, China).

Techniques: Inhibition, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: All-trans retinoic acid pretreatment of mesenchymal stem cells enhances the therapeutic effect on acute kidney injury

doi: 10.1186/s12964-024-01671-1

Figure Lengend Snippet: Schematic illustration of ATRA promoted HA production by MSCs and activated the PI3K/AKT pathway by binding to CD44 in renal tubular epithelial cells, thereby improving renal repair after AKI

Article Snippet: Human umbilical cord mesenchymal stem cells (hUC-MSCs) were acquired from the Beijing Zhongyuan Company (Beijing, China).

Techniques: Binding Assay

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Dose- and time-dependent effects of Fe 3 O 4 @Au nanoparticles on the viability of HPMSCs. Cells were incubated with increasing concentrations of Fe 3 O 4 @Au NPs (10–500 µg/mL). ( A ) The mitochondrial metabolic activity of HPMSCs was quantified via an MTT assay after 24 h, ( B ) 48 h, and ( C ) 72 h of incubation with Fe 3 O 4 @Au NPs, and ( D ) cell counting was performed via the trypan blue exclusion method on cells after 24 h, ( E ) 48 h, or ( F ) 72 h of incubation with Fe 3 O 4 @Au NPs. Data are expressed as the mean ± SEM of three independent experiments. *: p < 0.05, **: p < 0.01, ***: p < 0.001, and ****: p < 0.0001 as compared to control conditions.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Incubation, Activity Assay, MTT Assay, Cell Counting, Control

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Evaluation of the cellular uptake of Fe 3 O 4 @Au nanoparticles by HPMSCs. Cellular internalization was assessed using Prussian blue staining. The internalization of Fe 3 O 4 @Au NPs was determined on ( A ) control cells and ( B ) cells exposed to 100 µg/mL Fe 3 O 4 @Au NPs for 24 h, as well as ( C ) control cells and ( D ) cells exposed to 100 µg/mL Fe 3 O 4 @Au for 72 h. Blue staining is consistent with the intracellular uptake of Fe 3 O 4 @Au NPs by HPMSCs.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Staining, Control

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Effect of Fe 3 O 4 @Au nanoparticles on the necrosis and apoptosis markers of HPMSCs. ( A ) The cytotoxic effect of 100 µg/mL Fe 3 O 4 @Au NPs on cells was assessed using an LDH assay after 24 h and ( B ) 72 h of exposure to nanoparticles. The mean expression of genes encoding ( C ) caspase 3, ( D ) caspase 9, and ( E ) Bcl2 was obtained via RT-qPCR on HPMSCs incubated or not with Fe 3 O 4 @Au NPs for 24 h and 72 h. Data are indicated as the mean ± SEM of three independent experiments. *: p < 0.05, **: p < 0.01, and ****: p < 0.0001 as compared to the corresponding control conditions.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Lactate Dehydrogenase Assay, Expressing, Quantitative RT-PCR, Incubation, Control

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Effect of Fe 3 O 4 @Au nanoparticles on the redox status of HPMSCs. The mean normalized expression of genes encoding redox markers was determined by RT-qPCR. The expressions of genes coding for ( A ) Nox4, ( B ) HMOX-1, and ( C ) Nrf2 in HPMSCs were measured after 24 h and 72 h of exposure to 100 µg/mL Fe 3 O 4 @Au NPs. ( D ) ROS production was determined in HPMSCs exposed to 100 µg/mL Fe 3 O 4 @Au NPs for 2 h by using the DCFH-DA probe. Data are expressed as the mean ± SEM of three independent experiments. **: p < 0.01 and ****: p < 0.0001 as compared to the corresponding control conditions.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Effect of Fe 3 O 4 @Au nanoparticles on proinflammatory cytokines and chemokines secreted by HPMSCs. The proinflammatory response of HPMSCs was evaluated through ELISA. The levels of ( A ) TNF-α, ( B ) IL-1β, ( C ) IL-6, ( D ) CXCL8, and ( E ) CCL5 secreted by HPMSCs were quantified after 24 or 72 h of exposure to 100 µg/mL of Fe 3 O 4 @Au NPs. Data are presented as the mean ± SEM of three independent experiments. *: p < 0.05, **: p < 0.01, ***: p < 0.001, and ****: p < 0.0001 as compared to the corresponding control condition.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Cancers

Article Title: The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study †

doi: 10.3390/cancers16183239

Figure Lengend Snippet: Effect of Fe 3 O 4 @Au nanoparticles on the expression and production of protumorigenic factors by the HPMSCs. The gene expression and secreted levels of protumorigenic markers were measured on cells exposed to 100 µg/mL Fe 3 O 4 @Au nanoparticles for 24 and 72 h. First, the mean expression of genes encoding ( A ) PI3K, ( B ) AMPK, ( C ) PDGF, and ( D ) VEGF were determined via the use of RT-qPCR. Then, ( E ) the level of VEGF secreted by HPMSCs was quantified through an ELISA. Data are expressed as the mean ± SEM of three independent experiments. *: p < 0.05 and ****: p < 0.0001 as compared to the corresponding control conditions.

Article Snippet: Primary human pulmonary mesenchymal stem cells (HPMSCs) isolated from human lung tissue were purchased from the ScienCell research laboratory (Carlsbad, CA, USA) and were cultured in Modified Eagle’s Medium (MEM; PAN Biotech, Aidenbach, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS; PAN Biotech), 1 mM of sodium pyruvate (PAN Biotech), 0.5 μg/mL of amphotericin B (PAN Biotech), 2 mM of L-glutamine (Biochrom AG, Berlin, Germany), and 0.1 mg/mL penicillin–streptomycin (PAN Biotech).

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control