Neuromuscular junction in a microfluidic device

Abstract

Malfunctions at the site of neuromuscular junction (NMJ) of post-injuries or diseases are major barriers to recovery of function. The ability to efficiently derive motor neurons (MN) from embryonic stem cells has indicated promise toward the development of new therapies in increasing functional outcomes post injury. Recent advances in micro-technologies have provided advanced culture platforms allowing compartmentalization of sub-cellular components of neurons. In this study, we combined these advances in science and technology to develop a compartmentalized in vitro NMJ model. The developed NMJ system is between mouse embryonic stem cell (mESC)-derived MNs and c2c12 myotubes cultured in a compartmentalized polydimethylsiloxane (PDMS) microfluidic device. While some functional in vitro NMJ systems have been reported, this system would further contribute to research in NMJ-related diseases by providing a system to study the site of action of NMJ aimed at improving promoting better functional recovery.

Neuromuscular Activity Induces Paracrine Signaling and Triggers Axonal Regrowth after Injury in Microfluidic Lab-On-Chip Devices

“…Concerning PN-modelling, several methods were developed to generate contractile myotubes on lab-on-chip devices (i.e., [9][10][11]). But more relevantly, numerous protocols focused on mimicking PN networks (allowing neuromuscular junction (NMJ) formation) using muscle-derived cells (i.e, [12]), spinal cord slices (i.e., [13]), stem cells (i.e., [14,15]), and induced pluripotent stem cells (i.e., [16]), and in some of these, muscle activity was also analyzed (i.e., [11,[16][17][18]). However, none of these methods monitor the molecular events that occur in both muscle cells and MN after a PN axotomy.…”

Modelling functional human neuromuscular junctions in a differentially-perturbable microfluidic environment, validated through recombinant monosynaptic pseudotyped ΔG-rabies virus tracing

“…Recordings of electrical activity of the motor neurons were recorded on day 8,15,22,28,35,42,49,56, 63 and 70 days in vitro (DIV). Each recording was 4 minutes long at 10 000Hz, and was made at least 24 hours after media replacement.…”

“…Indeed, a number of studies have demonstrated the applicability of various bioengineering methods for creating muscle-neuron co-cultures, including functional NMJs in vitro. [15][16][17][18][19][20][21] In the present study, we model functional human NMJs by compartmentalized culture of iPS cell-derived MN pseudo-organoids and myotubes in a differentially perturbable multi-nodal microfluidic chip and demonstrate that NMJ function can be validated through monosynaptic retrograde tracing using a designer pseudotyped ΔG-rabies virus (RV).…”

Three‐Dimensional Models of the Human Brain Development and Diseases

“…Recent studies on mimicking neuromuscular junctions using micro-fluidic devices revealed that this technology could closely recapitulate ALS conditions in a dish. [79,80] …”