6 research outputs found
A Novel Functional In Vitro Model that Recapitulates Human Muscle Disorders
Here, we aim to address the increasing need for a suitable human muscle in vitro model in order to advance in the knowledge of muscle pathophysiology and test novel therapies for muscle disorders. Our model is based on a simple 2D culture method that yields highly mature human myotubes under optimized environmental conditions. Culture conditions that produced functional and contractile human myotubes with an extended lifetime consisted in extracellular matrix overlay and addition of several trophic factors to the differentiation medium. In this work, we describe the generation of suitable models of muscular dystrophies (limb-girdle muscular dystrophy type 2A—LGMD2A and Duchenne) by silencing expression of key proteins in these myotubes. Western blot and immunocytochemical analyses demonstrated similar features between our knockdown human myotubes and dystrophic muscles in vivo, which support the general validity of our cellular models. We also found that both dystrophic models present higher resting cytosolic Ca2+ levels than controls, which support a common underlying deficit in calcium homeostasis. This novel human in vitro system would allow for high-throughput screening of new treatments for these muscular dystrophies as well as for other neuromuscular disorders. In addition, our model could be used to advance in our understanding of human skeletal muscle pathophysiology
Defining and Evaluating a Core Genome Multilocus Sequence Typing Scheme for Whole-Genome Sequence-Based Typing of Listeria monocytogenes
HRM curve profiles of the ten most frequent <i>spa</i> types performed on a Rotorgene Q real-time instrument.
<p>t032, t022, t041, t2203, t001, t002, t003, t5712, t008, t190 double spiked with t030 and t003 genomic DNA.</p
Unspiked HRM curve profiles of the ten most frequent <i>spa</i> types.
<p>t032, t022, t041, t2203, t001, t002, t003, t5712, t008, t190 on an LC 480 real-time instrument.</p
Unspiked vs. spiked HRM curve profiles.
<p>Base line spa type t002. HRM melting curve profiles in blue show unspiked samples and melting curve profiles in green show double spiked samples derived from spa types t002, t003, t008, t011, t023, t030, t032, t037, t041, t068, 190, t919, t2689, and t3220 on an LC 480 real-time instrument.</p