rat mdcs  (Charles River Laboratories)

Journal: Frontiers in Pharmacology

Article Title: Achieving Acetylcholine Receptor Clustering in Tissue-Engineered Skeletal Muscle Constructs In vitro through a Materials-Directed Agrin Delivery Approach

doi: 10.3389/fphar.2016.00508

Figure Lengend Snippet: Dramatic impact of bioreactor preconditioning on AChR expression and organization in 3D tissue engineered constructs seeded with rat MDCs. (A–C) Bioreactor preconditioning in constructs with embedded agrin-presenting microspheres induces robust appearance large tracts of AChR clusters (red), indicating that mechanical stretch and agrin presentation may act synergistically. Very little AChR clustering and no large tracts of AChR clusters were observed for (D) unstretched (no bioreactor preconditioning) constructs with embedded agrin-presenting or (E) bioreactor preconditioned constructs with embedded BSA-presenting microspheres. Green color indicates fibrin construct (by autofluorescence) and the DAPI staining (blue) indicates an immature cell phenotype. (F) Shows the quantification of the bungarotoxin intensity as pixel intensity per unit area as determined by a blinded observer for three separate images ( n = 3) from a bioreactor experiment. Error bars in (F) indicate standard error of the mean and ∗ indicates P < 0.05 of stretched (bioreactor preconditioned) constructs compared to unstretched (static) agrin-presenting microspheres or stretched (bioreactor preconditioned) BSA-presenting beads. Scale bar represents 20 μm.

Article Snippet: Separately, rat MDCs were obtained from 4 to 6 week old male Lewis rats (Charles River, Raleigh, NC, USA) and were isolated as previously described ( ; ).

Techniques: Expressing, Construct, Staining

Journal: Frontiers in Pharmacology

Article Title: Achieving Acetylcholine Receptor Clustering in Tissue-Engineered Skeletal Muscle Constructs In vitro through a Materials-Directed Agrin Delivery Approach

doi: 10.3389/fphar.2016.00508

Figure Lengend Snippet: Representative examples of AChR clustering in a bioreactor-preconditioned agrin-presenting tissue engineered construct. Shown are rat MDCs (blue nuclei) seeded on the surface of fibrin scaffolds (green autofluorescence) carrying embedded SiO2 microspheres with covalently surface-bound agrin. Cells exhibited AChR clustering (red) after constructs were subjected to 5 days of cyclic mechanical strain. In contrast to the discrete geometry of AChR clusters observed under static culture with C2C12 cells (see Figures and ), AChRs in these rat MDCs cultured on bioreactor-conditioned agrin-presenting scaffolds clustered into extensive structures of complex geometry (A–D) more reminiscent of the appearance of native MEPs in vivo . By contrast, AChR clusters were never observed under static 3D culture conditions (compare to Figure ), indicating mechanical stretch and agrin presentation may act synergistically in the formation of mature MEP-like structures in vitro . (A–D) Represent selected magnified views from a 3D-projected confocal z-stack, and are shown in the context of the larger section ( E,F , compare to Figures ). Scale bars represent 20 μm.

Article Snippet: Separately, rat MDCs were obtained from 4 to 6 week old male Lewis rats (Charles River, Raleigh, NC, USA) and were isolated as previously described ( ; ).

Techniques: Construct, Cell Culture, In Vivo, In Vitro

Journal: Journal of biomedical materials research. Part A

Article Title: Quantitative in vivo Cytokine Analysis at Synthetic Biomaterial Implant Sites

doi: 10.1002/jbm.a.31939

Figure Lengend Snippet: Table I

Article Snippet: Multiplex Immunoassay A multiplex cytokine immunoassay containing a rat cytokine panel (IL-1β, IL-2, IL-4, IL-6, IL-10, GRO/KC, MDC-1, TNFα) was purchased from Lincoplex (Millipore, Billerica, MA).

Techniques: Multiplex Assay, Detection Assay