Journal: Cells

Article Title: A Novel Technique of Amniotic Membrane Preparation Mimicking Limbal Epithelial Crypts Enhances the Number of Progenitor Cells upon Expansion

doi: 10.3390/cells12050738

Figure Lengend Snippet: Differentiation marker expression profile of the ex vivo expanded hLESCs cultured on flat and crypt-like HAMs. (1) Polygonal and flat-like cells of the middle layers of cultures on flat HAMs stained positive for KRT3/12 antibody (1 A , D ). KRT3/12 was less present in hLESCs cultured on undulated (1 B , E ) and looped HAMs (1 C , F ). CX43 was present in most of the hLESC cultures, regardless of HAM formation (1 D – F ). (2) KRT3/12 and CX43 proteins were present throughout all cornea layers (2 A , D ), but they were absent in some cells of the basal limbal epithelium anteriorly (2 B , E ) and posteriorly (2 C , F ). Scale bars are the same for all images; hLESCs*—human limbal epithelial stem cell cultures.

Article Snippet: Blocking of non-specific binding sites with 5% Bovine Serum Albumin (BSA, A9418) dissolved in Dulbecco’s Phosphate Buffered Saline (DPBS, 14190-144, Thermo Fisher Scientific) was conducted for 20 min. Further, slides were stained with primary antibodies diluted in 1% BSA for 1 h. Slides were stained using antibodies for the following progenitor markers: tumor protein p63 alpha (p63α, rabbit polyclonal, 1:200 dilution, 4892S, Cell Signaling, Beverly, MA, USA), SRY-Box Transcription Factor 9 (SOX9, 82630, Cell Signaling, rabbit monoclonal, 1:200), quiescence marker: CCAAT/enhancer-binding protein delta (CEBPD, rabbit polyclonal, 1:200 dilution, ab198320, Abcam, Cambridge, UK), and proliferation marker Ki-67 (rabbit monoclonal, 1:200, RM-9106-S, Thermo Scientific) and the following differentiation markers: cytokeratin 3/12 (KRT3/12, mouse monoclonal, 1:100 dilution, 08691431, MP biomedicals, Santa Ana, CA, USA) and connexin-43 (CX43, rabbit polyclonal, 1:300, C6219).

Techniques: Marker, Expressing, Ex Vivo, Cell Culture, Staining

Journal: International Journal of Molecular Sciences

Article Title: Differentiation Effects of Platelet-Rich Plasma Concentrations on Synovial Fluid Mesenchymal Stem Cells from Pigs Cultivated in Alginate Complex Hydrogel

doi: 10.3390/ijms160818507

Figure Lengend Snippet: Evaluation of differentiation potential of SF-MSCs and bone marrow (BM)-MSCs by gene expressions. Gene expression profiles of SF-MSCs and BM-MSCs after osteogenic, chondrogenic, and adipogenic induction. Among them, type I collagen, osteocalcin, and osteopontin were for osteogenesis; type II collagen and aggrecan were for chondrogenesis; and peroxisome proliferator activated receptor γ 2 (PPAγ2) and adipocyte protein 2 (aP2) were for adipogenesis, respectively. Three SF-MSCs and BM-MSCs samples were performed for this experiment.

Article Snippet: Following permeabilization with 0.2% Triton X-100 (USB Corp., Cleveland, OH, USA) and blocking solution treatment (5% non-fat milk in PBS with 0.1% Triton X-100) for 30 min at room temperature, the slides were incubated with mouse anti-porcine monoclonal primary antibodies of type I collagen, type II collagen, and PPAγ2 (Merck Millipore, Darmstadt, Germany), applied for 1.5 h at room temperature.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Differentiation Effects of Platelet-Rich Plasma Concentrations on Synovial Fluid Mesenchymal Stem Cells from Pigs Cultivated in Alginate Complex Hydrogel

doi: 10.3390/ijms160818507

Figure Lengend Snippet: Evaluation of differentiation potential of SF-MSCs by histological staining and immunofluorescence staining. Histological staining ( A ) and immunofluorescence staining ( B ) of SF-MSCs after osteogenic (Alizarin red S stain and Type I collagen), chondrogenic (Alcain blue stain and Type II collagen), and adipogenic (Oil red O stain and PPAγ2) induction. Green: extracellular matrix and blue: cell nuclei.

Article Snippet: Following permeabilization with 0.2% Triton X-100 (USB Corp., Cleveland, OH, USA) and blocking solution treatment (5% non-fat milk in PBS with 0.1% Triton X-100) for 30 min at room temperature, the slides were incubated with mouse anti-porcine monoclonal primary antibodies of type I collagen, type II collagen, and PPAγ2 (Merck Millipore, Darmstadt, Germany), applied for 1.5 h at room temperature.

Techniques: Staining, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Differentiation Effects of Platelet-Rich Plasma Concentrations on Synovial Fluid Mesenchymal Stem Cells from Pigs Cultivated in Alginate Complex Hydrogel

doi: 10.3390/ijms160818507

Figure Lengend Snippet: Sequences of primers used in real-time PCR.

Article Snippet: Following permeabilization with 0.2% Triton X-100 (USB Corp., Cleveland, OH, USA) and blocking solution treatment (5% non-fat milk in PBS with 0.1% Triton X-100) for 30 min at room temperature, the slides were incubated with mouse anti-porcine monoclonal primary antibodies of type I collagen, type II collagen, and PPAγ2 (Merck Millipore, Darmstadt, Germany), applied for 1.5 h at room temperature.

Techniques:

Journal: PLoS ONE

Article Title: Bruton's tyrosine kinase inhibitor BMS-986142 in experimental models of rheumatoid arthritis enhances efficacy of agents representing clinical standard-of-care

doi: 10.1371/journal.pone.0181782

Figure Lengend Snippet: ( A ) Mean clinical scores over the course of the study, ( B ) mean clinical scores at the end of the study (day 46), ( C ) histological evaluation of the right hind paws, (D) plasma cells as measured by FACS analysis performed on spleens from 5 mice per group (3 mice in naïve group; non-immunized mice), (E) CD38 expression (MFI) on splenic CD138+B220low plasma cells, (F) anti-collagen II IgG titers, and ( G ) pharmacokinetics of BMS-986142 measured on the last day of the study with the data represented as time after the morning dose. The dashed line represents the IC50 value determined in vitro against BCR-stimulated CD69 expression on B cells in mouse whole blood. Data for B through F shown as mean ± SEM. * p < 0.05 versus vehicle group, # p < 0.05 versus either treatment alone, n = 10/group.

Article Snippet: A mixture of 4 monoclonal anti-mouse type II collagen antibodies (1 mg of each) was administered IP to female BALB/c mice (Harlan Laboratories; 8–10 weeks old) [ ].

Techniques: Clinical Proteomics, Expressing, Drug discovery, In Vitro