Journal: Biochemistry and Biophysics Reports

Article Title: The immunosuppressive capacity of human mesenchymal stromal cells derived from amnion and bone marrow

doi: 10.1016/j.bbrep.2016.07.019

Figure Lengend Snippet: The characteristic of cultured MSCs. (A) BM-MSCs at day 7. (B) Oil red-O staining of BM-MSCs at day 21. (C) ALP staining of BM-MSCs at day 14. (D) AM-MSCs at day 10. (E) Oil red-O staining of AM-MSCs at day 28. (F) ALP staining of AM-MSCs at day 28. (G) Flow cytometric analysis of surface marker expression. The data shown are representative of the three different experiments.

Article Snippet: For adipogenic differentiation, 3×10 4 MSCs from each source were cultured in adipogenic differentiation medium (Hyclone ® Advance STEM™, USA) with complete change of medium every 3 days.

Techniques: Cell Culture, Staining, Marker, Expressing

Journal: Biochemistry and Biophysics Reports

Article Title: The immunosuppressive capacity of human mesenchymal stromal cells derived from amnion and bone marrow

doi: 10.1016/j.bbrep.2016.07.019

Figure Lengend Snippet: Mean value of proliferative index of responder T cells co-cultured with stimulator T cells or PHA activated T cells in the presence or absence of MSCs. #, * p <0.05 significantly different compared to MNC+stimulator and MNC+PHA, respectively.

Article Snippet: For adipogenic differentiation, 3×10 4 MSCs from each source were cultured in adipogenic differentiation medium (Hyclone ® Advance STEM™, USA) with complete change of medium every 3 days.

Techniques: Cell Culture

Journal: Biochemistry and Biophysics Reports

Article Title: The immunosuppressive capacity of human mesenchymal stromal cells derived from amnion and bone marrow

doi: 10.1016/j.bbrep.2016.07.019

Figure Lengend Snippet: (A–C) Relative gene expression in MSCs after cultured with PHA activated T cells or IFN-γ. (D, E) Western blot analysis of IDO expression in MSCs after cultured with PHA activated T cells or IFN-γ. * ,# p <0.05 significantly different compared to BM-MSCs+MNC and AM-MSC+MNC, respectively.

Article Snippet: For adipogenic differentiation, 3×10 4 MSCs from each source were cultured in adipogenic differentiation medium (Hyclone ® Advance STEM™, USA) with complete change of medium every 3 days.

Techniques: Expressing, Cell Culture, Western Blot

Journal: Biochemistry and Biophysics Reports

Article Title: The immunosuppressive capacity of human mesenchymal stromal cells derived from amnion and bone marrow

doi: 10.1016/j.bbrep.2016.07.019

Figure Lengend Snippet: Mean value of proliferative index of PHA activated T cells co-cultured with MSCs in the presence or absence of 1-MT (A), Indomethacin (B), and l -NAME (C). *, # , $p <0.05 significantly different compared to MNC+PHA+BM-MSCs, MNC+PHA+AM-MSC and MNC+PHA+BM-MSCs+1MT, respectively.

Article Snippet: For adipogenic differentiation, 3×10 4 MSCs from each source were cultured in adipogenic differentiation medium (Hyclone ® Advance STEM™, USA) with complete change of medium every 3 days.

Techniques: Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Characterization of 3D Organotypic Culture of Mouse Adipose-Derived Stem Cells

doi: 10.3390/ijms25073931

Figure Lengend Snippet: Assessment of 3D organotypic ADSC differentiation potential. ( A ) Representative images of 3D organotypic ADSCs during adipogenic differentiation on the 14th day. The decrease in organoid size relative to the nondifferentiation control group is apparent. Scale bar size is 200 μm. ( B ) Oil Red O staining performed on the 14th day of adipogenic differentiation. Positive staining for Oil Red O is observed only in peripheral cells of the matrix. Scale bar size is 400 μm. ( C ) Relative mRNA expression levels of adipocyte markers LPL and adipoQ in 3D ADSCs compared to 2D ADSCs as a positive control. The expression of LPL and adipoQ mRNA in 3D ADSCs was not detected, in contrast to their expression in 2D ADSCs. The values are the means ± standard deviations ( n = 4).

Article Snippet: Adipogenic induction media (STEMCELL Technologies, Vancouver, Canada) were then added and replaced every 3 days.

Techniques: Control, Staining, Expressing, Positive Control

Journal: Journal of Tissue Engineering

Article Title: Therapeutic potential of adipose-derived mesenchymal stem cell exosomes in tissue-engineered bladders

doi: 10.1177/20417314211001545

Figure Lengend Snippet: Identification of Ad-MSCs (a–c) and AMEs (d–f): (a) characterization of the fourth generation of Ad-MSCs was performed by flow cytometry, and the Ad-MSC histogram revealed that after purification, the cells mostly expressed mesenchymal cell markers. Differentiation assays showed that Ad-MSCs could differentiate into adipocytes (b) and osteocytes (c), which were stained with Oil Red O and Von Kossa, respectively. Scale bar = 100 μm. (d) TEM of exosomes isolated from MSCM-cultured Ad-MSCs for 3 days with 10% exosome-depleted FBS. Scale bars = 100 nm. (e) NTA found that most of these vesicles ranged in diameter from 30 to 150 nm. (f) Western blotting was performed with Ad-MSCs (MSCs) and AMEs (Exos). The expression of CD9, CD63, TSG101, and calnexin was detected. Ad-MSCs: adipose-derived mesenchymal stem cells; AMEs: adipose-derived mesenchymal stem cell exosomes; NTA: nanoparticle tracking analysis; TEM: transmission electron microscopy.

Article Snippet: The medium was changed to mesenchymal stem cell adipogenic differentiation medium (MADM, ScienCell) and mesenchymal stem cell osteogenic differentiation medium (MODM, ScienCell) when the cells reached 70–80% confluency.

Techniques: Flow Cytometry, Purification, Staining, Isolation, Cell Culture, Western Blot, Expressing, Derivative Assay, Transmission Assay, Electron Microscopy

Journal: World Journal of Stem Cells

Article Title: CR6-interacting factor-1 contributes to osteoclastogenesis by inducing receptor activator of nuclear factor κB ligand after radiation

doi: 10.4252/wjsc.v12.i3.222

Figure Lengend Snippet: CR6-interacting factor-1 mediates adipogenesis and receptor activator of nuclear factor κB ligand secretion in adipocytes. A: Oil red O staining analysis of mouse bone marrow mesenchymal stem/stromal cells (BM-MSCs) after 21 d of adipogenic differentiation. Crif1 was knocked out in mouse BM-MSCs (BM-MSCs-KO), and knockout cells and controls were irradiated with 9 Gy of Co-60, and then treated with mouse mesenchymal stem cell adipogenic differentiation medium (Ad) to induce adipogenesis; B: The dye from oil red O staining was extracted using isopropanol, and the optical density at 510 nm was measured using Benchmark Plus; C: Western blot analysis of adipogenesis-related markers and transcription factors PPARγ and AP2 in mouse BM-MSCs after 21 d of adipogenic differentiation; D: Real-time quantitative polymerase chain reaction analysis of receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) mRNA expression in BM-MSCs and BM-MSCs-KO; E: RANKL/OPG ratio based on real-time quantitative polymerase chain reaction results; F: Enzyme linked immunosorbent assay analysis of RANKL protein levels in supernatant Ad; G: Enzyme linked immunosorbent assay analysis of OPG protein levels in supernatant Ad; H: RANKL/OPG ratio in supernatant Ad. aP < 0.05 vs control (BM-MSCs), bP < 0.01 vs control (BM-MSCs); dP < 0.01 between 9 Gy-BM-MSCs and 9 Gy-BM-MSCs-KO, and the bars represent the mean ± SD. BM-MSCs: Bone marrow mesenchymal stem/stromal cells; BM-MSCs-KO: Crif1 knockout mouse BM-MSCs; OPG: Osteoprotegerin; RANKL: Receptor activator of nuclear factor κB ligand.

Article Snippet: To induce adipogenesis, mouse BM-MSCs were seeded at a density of 2 × 10 4 cells per well in 6-well plates and cultured in mouse mesenchymal stem cell adipogenic differentiation medium (MUCMX-90031, Cyagen Biosciences).

Techniques: Staining, Knock-Out, Irradiation, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: World Journal of Stem Cells

Article Title: CR6-interacting factor-1 contributes to osteoclastogenesis by inducing receptor activator of nuclear factor κB ligand after radiation

doi: 10.4252/wjsc.v12.i3.222

Figure Lengend Snippet: CR6-interacting factor-1 inhibitors effectively suppress receptor activator of nuclear factor κB ligand secretion and adipogenesis. A: Enzyme linked immunosorbent assay (ELISA) analysis of receptor activator of nuclear factor κB ligand protein levels in the supernatant medium. Human bone marrow mesenchymal stem/stromal cells (H-BM-MSCs) were pretreated with five different compounds (25 µmol/L) followed by treatment with forskolin (25 µmol/L), and supernatant medium was collected for ELISA after 3 d; B: ELISA analysis of osteoprotegerin protein levels in supernatant medium; C: Receptor activator of nuclear factor κB ligand/osteoprotegerin ratio in supernatant medium; D: Oil red O staining analysis of H-BM-MSCs after 21 d of adipogenic differentiation. H-BM-MSCs were pretreated with five different compounds (25 µmol/L) followed by adipogenic induction; E: The dye from oil red O staining was extracted using isopropanol, and the optical density at 510 nm was measured using Benchmark Plus; F: Western blot analysis of cyclic adenosine monophosphate response element-binding protein phosphorylation levels. H-BM-MSCs were pretreated with five different compounds (25 µmol/L) followed by treatment with forskolin (25 µmol/L) and total protein lysates were extracted for cyclic adenosine monophosphate response element-binding protein phosphorylation detection after 1 h. aP < 0.05, bP < 0.01, and the bars represent the mean ± SD. OPG: Osteoprotegerin; RANKL: Receptor activator of nuclear factor κB ligand; CREB: Cyclic adenosine monophosphate response element-binding protein; H-BM-MSCs: Human bone marrow mesenchymal stem/stromal cells.

Article Snippet: To induce adipogenesis, mouse BM-MSCs were seeded at a density of 2 × 10 4 cells per well in 6-well plates and cultured in mouse mesenchymal stem cell adipogenic differentiation medium (MUCMX-90031, Cyagen Biosciences).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Binding Assay, Phospho-proteomics

Journal: NPJ Regenerative Medicine

Article Title: Personalized medicine for reconstruction of critical-size bone defects – a translational approach with customizable vascularized bone tissue

doi: 10.1038/s41536-021-00158-8

Figure Lengend Snippet: a EPC after uptake of acetylated LDL-DiI (red) and lectin staining (green). Counterstaining with DAPI (blue). Scale bar 50 µm. b CD34 immunofluorescence staining (green) of EPC. Counterstaining with DAPI (blue). Scale bar 50 µm. c Sprouting of EPC spheroids after 48 h incubation in a fibrin gel. Scale bar 50 µm. d mRNA expression of EPC compared to MSC (set to 1). e Oil Red O staining of MSC after adipogenic differentiation. Scale bar 100 µm. f Alcian Blue staining of MSC after chondrogenic differentiation. Scale bar 200 µm. g Alizarin Red staining of MSC after osteogenic differentiation. Scale bar 200 µm.

Article Snippet: For the adipogenic differentiation, MSC were cultivated in a differentiation medium consisting of MSC adipogenic differentiation basal medium, supplemented with FCS, glutamine, insulin, 3-isobutyl-1-methyl-xanthine (IBMX), dexamethasone, rosiglitazone, and penicillin-streptomycin; for chondrogenic differentiation MSC were cultivated in MSC chondrogenic differentiation basal medium, supplemented with FCS, dexamethasone, ascorbate, sodium selenite (ITS), proline, TGF-β3, and penicillin-streptomycin; for osteogenic differentiation MSC were cultivated in MSC osteogenic differentiation basal medium, supplemented with FCS, glutamine, ascorbate, β-glycerophosphate, dexamethasone, and penicillin-streptomycin according to the manufacturer’s recommendation (all from Pelobiotech GmbH, Planegg, Germany).

Techniques: Staining, Immunofluorescence, Incubation, Expressing