Abstract Details

Title Multielectrode array analysis of human iPSC-motor neuron maturation following co-culture with iPSC- spinal cord astrocytes

Topic Neuromuscular and Clinical Neurophysiology (EMG)

Presentation(s) P4 - Poster Session 4 (5:30 PM-6:30 PM)

Poster/Presentation
Number 12-031

Objective Develop a fully humanized co-culture platform of induced pluripotent stem cell-derived spinal cord astrocytes (hiPSC-A) and motor neurons (hiPSC-MN) for multi-electrode array (MEA) recording.

Background MEA has been previously utilized in electrophysiological recordings of rodent MNs and hiPSC-cortical neurons. However, there is no literature on hiPSC-A influences on hiPSC-MN electrophysiological properties.

Design/Methods Through the critical steps of caudalization and ventralization, we generated spinal cord hiPSC-A and -MNs. We compared hiPSC-MN monocultures to hiPSC-MN and hiPSC-A co-cultures and investigated astrocyte variables that could influence MN maturation. MEA recordings were performed weekly over a four-week period, to obtain electrophysiological parameters including spike and burst rate and the percentage of spiking and bursting electrodes. Immunocytochemistry was used to analyze astrocyte and neuronal populations. We tested the effect of neurotransmitter agonists/antagonists, including kainate, CNQX and bicuculline.

Results The culture of hiPSC-MN alone resulted in large aggregates of cells with delayed electrophysiological maturation. The addition of hiPSC-A resulted in reduced neuronal aggregation accompanied by the accelerated electrophysiological maturation of hiPSC-MN across all time points examined. These electrophysiological measures following hiPSC-A/hiPSC-MN co-culture were mirrored morphologically by increased diameter of neuronal processes, mean soma size, and complexity of neuronal connections. We also found that hiPSC-MN and hiPSC-neurons expressed appropriate neurotransmitter receptors which were modulated by known agonist/antagonists. Using several lines of hiPSC-A, we also found that the electrophysiological properties are consistent across lines. When compared with rodent astrocytes, hiPSC-A also appear to induce a more rapid maturation of hiPSC-MN both morphologically and electrophysiologically.

Conclusions We demonstrate that this humanized co-culture method allows for the long-term, non-invasive, serial recording of spinal cord neuronal populations, and for the electrophysiological analysis of the effects of pharmacologic manipulation of these cultures. This platform will be of use for studying astrocyte/neuron interactions in the context of motor neuron disease.

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
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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