Journal: PLoS ONE

Article Title: Cell Population Kinetics of Collagen Scaffolds in Ex Vivo Oral Wound Repair

doi: 10.1371/journal.pone.0112680

Figure Lengend Snippet: The ex vivo wound healing model is shown in top view and cross-sectional view (A). The bottom of the well was covered with an acellular growth factor reduced basal membrane extract (BME). The scaffold (sponge type scaffold composed of cross-linked collagen or the gel type scaffold composed of rat collagen I) in the center was surrounded by a BME-gel containing 800,000 hGFs/ml and covered with an acellular BME gel and Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 1% fetal bovine serum (FBS) and antibiotics. Both scaffold and gel were tested with and without the addition of platelet-derived growth factor-BB (PDGF). Fluorescence images were taken to assess DiI-labeled cells within the scaffold over time. To assess the metabolic activity and gene expression dynamics, the scaffolds including the “active zone” (dashed line) were subjected to MTT tests, gene array analysis, and quantitative PCR. (B) Representative scanning electron microscopy images depict the morphology of the collagen gel, collagen scaffold at 400×, and 800× magnification. The white bar represents 100 µm.

Article Snippet: 48 well plates were first layered with 50 μL of 12 mg/ml growth factor reduced basal membrane extract (BME; CULTREX, TREVIGEN, Gaithersburg, MD, USA) without cells.

Techniques: Ex Vivo, Membrane, Modification, Derivative Assay, Fluorescence, Labeling, Activity Assay, Gene Expression, Real-time Polymerase Chain Reaction, Electron Microscopy

Journal: Journal of Virology

Article Title: Development of a Primary Human Cell Model for the Study of Human Cytomegalovirus Replication and Spread within Salivary Epithelium

doi: 10.1128/JVI.01608-18

Figure Lengend Snippet: Establishment of salispheres and primary epithelial lines from human salivary gland. (A) Freshly isolated salivary gland tissues (top) were digested with collagenase-dispase, and single-cell suspensions were then cultured on Cultrex PathClear basement membrane extract (BME) gels, where multicellular salispheres developed within 72 to 96 h (middle). Salispheres were then dissociated, counted, and replated onto Cultrex PathClear BME gels as in the middle panel or plated onto plastic culture dishes to facilitate HCMV infection experiments (bottom). (B) Salispheres were used for mRNA preparation and amylase 1 and hypoxanthine-guanine phosphotransferase (HPRT) expression. (C) Salispheres were then used for more-detailed analyses of salivary gland-specific gene expression. Taken together, the results indicate that the salispheres grown in vitro exhibit gene expression profiles similar to those of salivary tissue in vivo.

Article Snippet: Following digestion and lysis of red blood cells, the salivary cells were plated on Cultrex PathClear basement membrane extract (BME) gels (R&D Systems) and cultured in BEGM growth medium (containing the BEGM bullet kit and 4% charcoal-stripped fetal bovine serum [FBS]), provided by Lonza.

Techniques: Isolation, Cell Culture, Membrane, Infection, Expressing, Gene Expression, In Vitro, In Vivo