Journal: Frontiers in Cell and Developmental Biology

Article Title: Conditions Mimicking the Cancer Microenvironment Modulate the Functional Outcome of Human Chorionic Villus Mesenchymal Stem/Stromal Cells in vitro

doi: 10.3389/fcell.2021.650125

Figure Lengend Snippet: Effect of CM-MDA231 on CV-MSCs expression of Adhesion molecules. Flow cytometry analysis of the adhesion molecules performed on preconditioned, in-treatment, and untreated CV-MSCs showed that there was significant increase in the expression levels for VCAM, ICAM1 and PECAM, after preconditioning them for 72 h (A–C) , but no changes in the expression levels were observed in cells preconditioned for 24 h, in-treatment and untreated controls. No significant changes in expression levels was observed for E-Cadherin (D) between the preconditioned, in-treatment and untreated controls (Panel 1). The data obtained by FACS from five independent experiments was quantified. The average is presented as bar diagrams in panel 2. VCAM is represented as (A) , ICAM1 as (B) , PECAM as (C) and E-Cadherin as (D) in panel 2, respectively. Bars represent standard errors * P ≤ 0.05.

Article Snippet: GAPDH and a panel of fluorescent-labeled antibodies (VCAM (cat#FAB5649P), ICAM (cat#BBA20), PECAM (cat#FAB3567P), E-Cadherin (cat#FAB18381P), Integrin α5 (cat#FAB1864P), Integrin-M (cat#FAB16991P), and EpCAM (cat#FAB9601P), used for flow cytometry were purchased from R&D Systems, MN, United States. p53 (cat#2527), pRb (S807) (cat#8516), pChk2 (T68) (cat#2661) and β-Actin (cat#3700 and cat#8457) antibodies for immunoblotting were purchased from Cell Signaling Technologies, MA, United States.

Techniques: Expressing, Flow Cytometry

Journal: Scientific Reports

Article Title: A novel strategy to engineer pre-vascularized 3-dimensional skin substitutes to achieve efficient, functional engraftment

doi: 10.1038/s41598-019-44113-6

Figure Lengend Snippet: Interconnected vascular network formation in 3D skin substitutes 7 days after epidermal differentiation. ( a – c ) HUVEC were seeded at 0.1 to 1 × 10 5 cells per culture insert; FN-G-coated NHDF content (1 × 10 7 cells) remained constant. FN-G-coated NHDF and HUVEC were cocultured at 1,000:1, 500:1, or 100:1, and then with HEKn (1 × 10 6 cells). ( a ) Whole-mount CD31 immunofluorescence analysis revealed that HUVEC developed branching vessels within the dermal layer. The data are representative of 2 independent experiments (n = 6/condition). ( b , c ) Quantification of vascular structures to determine the vessel density ( b ) and branching index (branching points/unit area) ( c ). The data are the mean ± SD (n = 6). * p < 0.05 and ** p < 0.01 determined by one-way ANOVA with Tukey’s multiple comparison post hoc test.

Article Snippet: The following primary antibodies were used for immunostaining of: a human–specific CD31 (1:200, NBP2-15202, clone C31.3; Novus Biologicals, Centennial, CO, USA), a mouse–specific CD31 (1:100, 14-0311-82, clone 390; Invitrogen), HLA-Class I ABC (1:2,500, ab70328, clone: EMR8-5; Abcam), and laminin 5 (1:200; ab14509, Abcam).

Techniques: Immunofluorescence, Comparison

Journal: Scientific Reports

Article Title: A novel strategy to engineer pre-vascularized 3-dimensional skin substitutes to achieve efficient, functional engraftment

doi: 10.1038/s41598-019-44113-6

Figure Lengend Snippet: In vitro evaluation of the therapeutic potential of pre-vascularized 3D skin substitutes. ( a – e ) HUVEC (0.2 × 10 5 cells/insert) mixed with FN-G-coated NHDF were cocultured, subsequently covered with HEKn, then cultured for up to an additional 7 days. ( a ) Macroscopic view of the construct in the culture insert. Scale bar: 10 mm. ( b – e ) Histological and immunohistochemical staining with hematoxylin and eosin ( b ), Masson’s trichrome ( c ), anti-laminin 5 ( d , arrows = basement membrane), and anti-CD31 (e, arrows = CD31 + blood vessel). Scale bars: 500 μm ( b ), 100 μm ( c – e ).

Article Snippet: The following primary antibodies were used for immunostaining of: a human–specific CD31 (1:200, NBP2-15202, clone C31.3; Novus Biologicals, Centennial, CO, USA), a mouse–specific CD31 (1:100, 14-0311-82, clone 390; Invitrogen), HLA-Class I ABC (1:2,500, ab70328, clone: EMR8-5; Abcam), and laminin 5 (1:200; ab14509, Abcam).

Techniques: In Vitro, Cell Culture, Construct, Immunohistochemical staining, Staining, Membrane

Journal: Scientific Reports

Article Title: A novel strategy to engineer pre-vascularized 3-dimensional skin substitutes to achieve efficient, functional engraftment

doi: 10.1038/s41598-019-44113-6

Figure Lengend Snippet: Quantification of wound vasculature in vivo . ( a ) Immunohistochemical detection of mouse-specific CD31 (mCD31) and human-specific CD31 (hCD31) at 7 and 14 days after grafting. Dashed lines indicate the skin substitute–host interface. Scale bars: 100 μm. ( b ) Quantification of CD31 + blood vessels in the dermal areas of grafts. The data are the mean ± SD (n = 5). ** p < 0.01 compared with the non-vascularized control (unpaired Student’s t -test). ( c ) Immunofluorescence image stained for mouse-CD31 (red) and human-CD31 (green) of pre-vascularized substitute-treated wounds at day 14 after grafting. Nuclei were stained with DAPI (blue). White arrowheads indicate human–mouse chimeric vessel. Scale bars: 50 μm.

Article Snippet: The following primary antibodies were used for immunostaining of: a human–specific CD31 (1:200, NBP2-15202, clone C31.3; Novus Biologicals, Centennial, CO, USA), a mouse–specific CD31 (1:100, 14-0311-82, clone 390; Invitrogen), HLA-Class I ABC (1:2,500, ab70328, clone: EMR8-5; Abcam), and laminin 5 (1:200; ab14509, Abcam).

Techniques: In Vivo, Immunohistochemical staining, Control, Immunofluorescence, Staining