Devices and methods for preparing in vitro models of neuromuscular junctions
Inventors
GEORGE, Subin Mac • Kuyltjes, Lindsay Morgan • Magdesian, Margaret Haiganouch
Assignees
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Publication Number
US-12173266-B1
Publication Date
2024-12-24
Expiration Date
Abstract
There are provided a cell culture device and associated method for preparing an in vitro model of a neuromuscular junction (NMJ). The cell culture device can include a cell culture layer that includes a neuronal cells channel for receiving neuronal cells therein, and a neuromuscular junction chamber in fluid communication with the neuronal cells channel to receive muscle cells therein to enable co-culture of the neuronal cells with the muscle cells and form the NMJ. The neuronal cells channel includes a first portion having a first portion cross-section and a second portion having a second portion cross-section that is smaller than the first portion cross-section and provided downstream of the first portion to retain neuronal cells. The neuromuscular junction chamber can optionally include pillars for culturing muscle cells therebetween. The in vitro model can be used for analysing a biological material from a patient or diagnose a condition, for instance.
Core Innovation
The invention relates to an in vitro cell culture system for a neuromuscular junction that includes a neuronal cells inlet and a neuronal cells channel. The neuronal cells channel has a first portion sized to retain neural cell bodies and a downstream second portion with a reduced cross-section that prevents entry of cell bodies while allowing axons to reach an NMJ chamber for neuronal-muscle co-culture.
The in vitro system includes a neuromuscular junction chamber configured for co-culture between axons and muscle cells, and it may include muscle anchoring pillars or anchoring locations to position or anchor muscle in the chamber. The geometry of the neuronal channel and the downstream reduced cross-section, including an hourglass section, function to bias the relationship between retained cell bodies and axonal extension toward the NMJ chamber.
The invention further describes optional enhancements for the cell culture system, including a gel seeding inlet configured to seed or embed muscle within a matrix, and microchannels or longitudinally extending support structures in the neuronal channel to bias axonal growth and stabilize neurospheres. The NMJ chamber may further include an electrode layer or multi-electrode array and detectors to provide electrical stimulation and recording for assessing functionality parameters including contraction force, duration, viability, and electrical signals, including responses to test substances.
Claims Coverage
The claims coverage identifies the in vitro cell culture system for a neuromuscular junction and its associated chamber and channel features. One inventive feature is explicitly reflected in the claimed geometry and functional separation of cell bodies and axons.
Neuronal cells channel with reduced cross-section
A neuronal cells channel has a first portion sized to retain neural cell bodies and a downstream second portion with a reduced cross-section that prevents entry of cell bodies while allowing axons to reach an NMJ chamber.
Neuromuscular junction chamber for co-culture
An NMJ chamber is configured for co-culture between axons and muscle cells, with muscle anchoring pillars or anchoring locations to position or anchor muscle in the chamber.
Channel geometry to bias axonal extension
The geometry of the neuronal channel, including an hourglass section, biases the relationship between retained cell bodies and axonal extension toward the NMJ chamber.
Optional seeding and sensing features
Optional features include a gel seeding inlet to seed or embed muscle within a matrix, microchannels or longitudinally extending support structures to bias axonal growth and stabilize neurospheres, and an electrode layer or multi-electrode array and detectors for electrical stimulation and recording.
The claims collectively cover the channel geometry, the NMJ chamber for neuronal-muscle co-culture, and the optional seeding, support, and sensing features.
Stated Advantages
Prevents entry of neural cell bodies into the downstream neuronal channel while allowing axons to reach an NMJ chamber for neuronal-muscle co-culture.
Anchors muscle in the NMJ chamber using muscle anchoring pillars or anchoring locations.
Biases axonal growth and stabilizes neurospheres using microchannels or longitudinally extending support structures.
Enables embedding muscle in a matrix using a gel seeding inlet.
Enables electrical stimulation and recording to measure contraction force, duration, viability, and electrical signals.
Supports analysis of patient-derived biological materials for diagnosing a condition and assessing the effects of a test substance, including Botulinum toxin.
Documented Applications
Analyzing patient-derived biological materials for diagnosis of a condition.
Assessing drug or toxin effects of a test substance, including Botulinum toxin, using the NMJ in vitro model.
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