## 9 Tissue Engineering  
Differentiation and Characterization of hiPSC-Cortical Neurons and Their Application to Drug Evaluation in CNS Disease Models  
[PERSON] \({}^{1}\), [PERSON] \({}^{1}\), [PERSON] \({}^{1}\), [PERSON] \({}^{1}\), [PERSON] \({}^{2}\), [PERSON] \({}^{2}\), [PERSON] \({}^{2}\), [PERSON] \({}^{2}\) and [PERSON] \({}^{1,2}\)  
\({}^{1}\) Nanoscience Technology Center, University of Central Florida, 12424 Research Parkway, Suite 400, Orlando, FL 32826, USA  
\({}^{2}\) Hesperos, Inc. 3459 Progress Dr. Orlando, FL 32826, USA  
The differentiation of functional cortical neurons from human induced pluripotent stem cells (hiPSCs) in vitro easily lends itself to a serum-free, drug delivery platform advantageous for testing novel chemicals for safety and efficacy in disease treatment. Initially, cortical neuron cultures were characterized morphologically by phase microscopy and immunocyt and functionally by patch-clamp electrophysiology. Specifically, the expression of neuronal markers and neuronal activity increased throughout maturation. On day 0 of maturation, 50 percent of the culture expressed layer V cortical neuron marker ctip2 and neuronal marker beta-III tubulin and displayed spontaneous and repetitive firing through whole-cell patch clamp. By day 28 of maturation, 90 percent of the culture expressed the aforementioned markers and displayed electrical activity. Subsequently, neurons were cultured on multi-electrode arrays (MEAs) to determine the effects of chemicals on neural circuit physiology for modeling brain disease phenotypes. Long-term potentiation (LTP) was induced via a high-frequency stimulation (HFS), which was subsequently abolished following the dosage with Na\({}^{+}\) channel blocker lidocaine. In this system, we also tested GABA\({}_{\text{A}}\) receptor antagonists and agonists as chemical convulsants or anti-convulsants, respectively. GABA\({}_{\text{A}}\) receptor antagonist administration enhanced spontaneous activity mimicking an epileptic phenotype that further increased upon electrical stimulation, while GABA\({}_{\text{A}}\) receptor agonist administration quieted spontaneous activity. The versatility of this model lies in its ability to present an array of brain diseases characterized by functional brain deficits. Chemicals affecting receptor binding can be added to either enhance or inhibit neuronal activity. This serum-free, hiPSC cortical neuron model establishes a platform for the evaluation of neuron activity as well as a platform for drug testing in vitro. Cantilever-Based Analysis of Skeletal Muscle Force Output from ALS Patient-Derived iPSCs for Research and Drug Discovery  
[PERSON], [PERSON] and [PERSON]  
Hickman Hybrid Systems Laboratory, NanoScience Technology Center, University of Central Florida, Orlando, FL 32826, USA  
Amyotrophic lateral sclerosis (ALS) is a debilitating adult-onset disease, characterized by motoneuron degeneration and muscle wasting. It is currently incurable, and leads to death within 1-3 years of diagnosis. The development of curative treatments has been challenging, due to the limited capability of current disease models in predicting therapeutic outcomes in patients. Moreover, the multi-faceted nature of ALS poses major challenges in elucidating the disease's spatiotemporal progression. This necessitates the development of human-based models, capable of recapitulating disease conditions to help hasten the currently lengthy and expensive drug discovery process. Disruption of the neuromuscular junction (NMJ) is an established early event in ALS pathology. However, its cause is undetermined. Recent reports indicate that skeletal muscle pathology precedes NMJ dysregulation, motoneuron degeneration and can induce phenotypic ALS symptoms. Yet, the role of dysfunctional muscle in ALS onset is debatable, due to limited studies on the subject. Here, we describe the development of an in vitro phenotypic skeletal muscle model derived fromALS patient induced pluripotent stem cells (ALS-iPSCs).