Archived record · Edyssy 2014

An In Vitro Model Of Stem Cell Innervation Of Myotubes

Agency

NIHNational Institutes of Health

Principal investigator

James Hickman

Institution

University of Central Florida

Amount listed

$400,932

Period

Feb. 1, 2011 to Jan. 31, 2012

Abstract

DESCRIPTION (provided by applicant): A long-term goal of the research in our laboratory is to re-create the components and compartments of mammalian systems, and in particular human systems, to engineer functional hybrid models. The initial period for this Bioengineering Research Grant (BRG) was to investigate the motoneuron to muscle segment of the reflex arc to understand in vitro what are possible outcomes when a human stem cell is implanted in a rat model and then apply it to improve the outcomes in an in vivo system. We have made a number of breakthroughs with this model system that move us closer to the goal of creating a functional in vitro model of the reflex arc: 1) We have cultivated embryonic and adult rat motoneurons as well as motoneurons derived from human fetal stem cells (hFSC). 2) We have demonstrated neuromuscular junction formation between these rat and human derived motoneurons with myotubes derived from both rat and human adult satellite cells. However, we have also created mature myotubes that exhibit sarcomere formation, express myosin heavy chain, have established the EC coupling apparatus as well as Type I and Type II muscle isoforms. We have also differentiated the myotubes into intrafusal fibers and integrated then with the MEMS constructs and demonstrated that these are functional mechanoreceptor synapses. 3) Glial cells have been incorporated into our model system and we have demonstrated Schwann cell myelination of motoneurons, with complete Node of Ranvier formation, and derived pure populations of MBP (myelin basic protein) positive oligodendrocytes from human mesenchymal stem cells. 4) We have successfully demonstrated in vivo integration of the differentiated hFSC derived motoneurons into an adult rat spinal cord injury model and shown significant locomotor recovery as accessed via the BBB test scoring system. This renewal BRG application seeks to complete the development of the entire reflex arc in vitro using primary rat cells and then with cells derived from human stem cells. We will also initiate experiments to increase the throughput of first the motoneurons to muscle segment of the stretch reflex are and then for the entire reflex arc. The Hypothesis is that we can recreate a functional in vitro model of the reflex arc with primary rat cells and achieve the same result with cells derived from human stem cells that will be a prototype for the next generation screens for drug discovery and toxicology. The continuation of the grant will enable advances for 1) the creation of a human cell based reflex arc in a defined, serum-free environment and 2) the development of new technology for utilization in the next level of Systems Biology and high-information content screens. Our expertise in surface chemistry, Micro-ElectroMechanical Systems (MEMS) fabrication and developmental/cellular/molecular biology has now allowed the construction of complex functional neuronal systems in vitro. Our team possesses the necessary expertise in all aspects of this project, as well as the facilities to effectively and successfully perform the research.

PUBLIC HEALTH RELEVANCE: The goal of this project is to engineer a system to model one of the most fundamental motor circuits in the human body, the spinal reflex arc. We will use nanotechnology and microelectronics in combination with biomedical engineering techniques to build this hybrid biological/non-biological system. Potential benefits include prevention, diagnosis, and treatment of developmental abnormalities in the spinal cord, rehabilitation of chronic neurological/muscle disorders and accompanying pain, and new strategies for prosthetic and orthotic design and evaluation. Public Health Relevance Statement: The goal of this project is to engineer a system to model one of the most fundamental motor circuits in the human body, the spinal reflex arc. We will use nanotechnology and microelectronics in combination with biomedical engineering techniques to build this hybrid biological/non-biological system. Potential benefits include prevention, diagnosis, and treatment of developmental abnormalities in the spinal cord, rehabilitation of chronic neurological/muscle disorders and accompanying pain, and new strategies for prosthetic and orthotic design and evaluation. The goal of this project is to engineer a system to model one of the most fundamental motor circuits in the human body, the spinal reflex arc. We will use nanotechnology and microelectronics in combination with biomedical engineering techniques to build this hybrid biological/non-biological system. Potential benefits include prevention, diagnosis, and treatment of developmental abnormalities in the spinal cord, rehabilitation of chronic neurological/muscle disorders and accompanying pain, and new strategies for prosthetic and orthotic design and evaluation. Adult;Afferent Neurons;Axon;base;Biocompatible Materials;Biological;Biological Assay;Biological Models;Biology;Biomedical Engineering;cell type;Cells;Chemistry;Chronic;Complex;Coupling;Data;design;Development;Devices;Diagnosis;drug discovery;Embryo;Engineering;Environment;Evaluation;Exhibits;fetal stem cell;Fiber;Funding;Goals;Grant;Hippocampus (Brain);Human;Human body;human stem cells;Hybrids;Implant;improved;In Vitro;in vitro Model;in vivo;information processing;innovation;Journals;Laboratories;Legal patent;Locomotor Recovery;Manuscripts;Mechanoreceptors;Mesenchymal Stem Cells;Microelectrodes;Modeling;Molecular and Cellular Biology;Motor;Motor Neurons;Muscle;Muscle Fibers;Myelin Basic Proteins;myelination;Myopathy;Myosin Heavy Chains;Nanotechnology;Natural regeneration;Neonatal;nerve supply;Nervous system structure;Neuroglia;Neurologic;Neuromuscular Junction;Neurons;Neurotransmitters;new technology;next generation;Oligodendroglia;Orthotic Medical Specialty;Outcome;Pain;Population;Preparation;Prevention;Production;Progress Reports;Prosthesis;Protein Isoforms;prototype;Publications;Ranvier's Nodes;Rattus;Reflex action;Rehabilitation therapy;Research;Research Project Grants;research study;Sarcomeres;satellite cell;Schwann Cells;Screening procedure;Serum;solid state;Spinal Cord;Spinal cord damage;Spinal cord injury;spinal reflex;Stem cells;stretch reflex;Structure;Surface;symposium;Synapses;synaptic function;System;Systems Biology;Techniques;Technology;Testing;Toxicology;Work

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The record names James Hickman at University of Central Florida as principal investigator. When a professor holds an award like this, part of the budget usually goes to people: graduate research assistants, a postdoc or a technician. Shorter awards are often one year of a renewing project, so check whether it was continued. Look up the investigator’s current awards before you write; a 2014 record is a lead, not a promise. Before you do, read up on a first email to a lab like this.

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