Abstract Details

Title Modeling the Human Corticospinal Tract-on-a-chip with Regionally-specific hiPSC-derived Neurons and Astrocytes

Topic Neuromuscular and Clinical Neurophysiology (EMG)

Presentation(s) S46 - Neuromuscular and Clinical Neurophysiology (EMG): Motor Neuron/Charcot Marie Tooth (4:54 PM-5:06 PM)

Poster/Presentation
Number 008

Background The co-culture of regionally specific hiPSC-N/–A has been used to model the neural circuits of the cortex and the spinal cord. The relative immaturity of these cells and their networks remains a major limitation for modeling Amyotrophic Lateral Sclerosis (ALS), whose pathological abnormalities are age-dependent and occur within the CST.

Objective Develop a microfluidic system to co-culture human pluripotent stem cells (hiPSC)-derived neurons (hiPSC-N) and astrocytes (hiPSC-A) displaying region-specific phenotypes (i.e. cortical and spinal cord) and model the connections of the human corticospinal tract (CST).

Design/Methods We adapted two previously published hiPSC-based protocols to generate hiPSC-N and -A with regionally-specific phenotypes, i.e. ChAT+ spinal motor neurons (hiPSC-SMN), CTIP2+ layer V corticospinal motor neurons (hiPSC-CSMN), “spinal cord” astrocytes (hiPSC-scA) as shown by immunoreactivity to HOXB4 and “cortical” hiPSC-A (hiPSC-cA) as indicated by immunoreactivity for CD51 and a lack of HOXB4 expression. We then co-cultured cortical and spinal cord hiPSC-N and –A in two separated compartments (hiPSC-CSMN/hiPSC-cA and hiPSC-SMN/hiPSC-scA, respectively) of a 3 chamber microfluidic chip.

Results

We observed the emergence of axonal projections between the “cortical” and “spinal cord” compartments of the microfluidic system which model the connections of the corticospinal tract. Interestingly, the connections between cortical and spinal neurons appeared earlier and were more robust when compared to those observed for only spinal or only cortical populations. We then used multielectrode array recording (MEA) and recorded electrophysiological activities from both cortical and spinal cord neuronal populations.

Finally, when primary mouse myotubes were plated in the third compartment of the microfluidic system, we observed the formation of neuromuscular junctions as indicated by co-localization of SV2/ αBTX.

Conclusions We develop a hiPSC- and microfluidic-based system to model in vitro the organization of the CST. This platform will be of use for studying network pathology in the context of ALS.

Authors/Disclosures
Arens Taga, MD (Johns Hopkins University) PRESENTER	Dr. Taga has nothing to disclose.
	No disclosure on file
	No disclosure on file
Christa Habela, MD (Johns Hopkins University)	The institution of Dr. Habela has received research support from NINDS. Dr. Habela has received publishing royalties from a publication relating to health care.
	No disclosure on file
	No disclosure on file
Arun Venkatesan, MD, PhD (Johns Hopkins Hospital)	Dr. Venkatesan has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Janssen Pharmaceuticals. The institution of Dr. Venkatesan has received research support from NIH. The institution of Dr. Venkatesan has received research support from MSRCF. The institution of Dr. Venkatesan has received research support from U.S. DOD.
Nicholas J. Maragakis, MD, FAAN (Johns Hopkins University School of Medicine)	Dr. Maragakis has received personal compensation in the range of $500-$4,999 for serving as a Consultant for UptoDate. Dr. Maragakis has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Apellis Pharmaceuticals. Dr. Maragakis has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Amylyx. An immediate family member of Dr. Maragakis has received personal compensation in the range of $10,000-$49,999 for serving as a Consultant for National Basketball Association. Dr. Maragakis has received personal compensation in the range of $0-$499 for serving on a Scientific Advisory or Data Safety Monitoring board for Akava. An immediate family member of Dr. Maragakis has received personal compensation in the range of $0-$499 for serving as an officer or member of the Board of Directors for Johns Hopkins Howard County Medical Center. Dr. Maragakis has stock in Akava. The institution of Dr. Maragakis has received research support from NIH/NINDS. The institution of Dr. Maragakis has received research support from Department of Defense. The institution of Dr. Maragakis has received research support from Maryland Stem Cell Research Fund. Dr. Maragakis has received intellectual property interests from a discovery or technology relating to health care.

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