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: World Journal of Gastroenterology : WJG

Article Title: On the origin of cardiac mucosa: A histological and immunohistoc-hemical study of cytokeratin expression patterns in the developing esophagogastric junction region and stomach

doi: 10.3748/wjg.v11.i29.4490

Figure Lengend Snippet: Tubular esophagus. A: Esophageal ciliated epithelium at 20-wk GA. The immunostaining is for CK5 and is more intense in the basal and intermediate cell layers than in the superficial cell layer (OM ×400); B: Esophageal squamous epithelium in a 3-wk-old neonate. The immunostaining is for CK13 and is absent in the basal cell layer and diffuse moderate to strong in the intermediate and superficial cell layers (OM ×200); C: Esophageal simple columnar epithelium at 20-wk GA. To the left: parietal cells with roughly triangular shape and highly acidophilic cytoplasm (ESP, arrowhead). To the right: no discernible parietal cells are present (ESN, H&E, OM ×400); D: Esophageal simple columnar epithelium at 20-wk GA. The immunostaining is for CK7 and shows moderate positivity at the mucosal surface and deep in the glands (arrowhead and arrow, respectively, OM ×200); E: Esophageal simple columnar epithelium at 20-wk GA (same case as in Figure ​Figure3D).3D). The immunostaining is for CK20 and shows patchy positivity of the superficial part of the mucosa (surface and pit epithelium: arrowhead and arrow, respectively, OM ×200).

Article Snippet: Murine mAbs against human CK7, 18, 19, and 20 were purchased from DakoCytomation (Glostrup, Denmark); mAbs against CK5, 8, and 13 were purchased from Novocastra Laboratories (Newcastle upon Tyne, UK, Table ).

Techniques: Immunostaining

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: