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Harvesting, Embedding, and Culturing Dorsal Root Ganglia in Multi-compartment Devices to Study Peripheral Neuronal
Benjamin M Gane1, Sydney M Caparaso1, Fei San Lee1
1Department of Biological Systems Engineering, University of Nebraska-Lincoln.
Journal of Visualized Experiments : Jove
|July 15, 2024
Summary
A novel in vitro model using intact dorsal root ganglia (DRGs) allows studying neural-immune cell interactions. This platform enhances pain research by providing a more accurate preclinical screening tool for potential pain treatments.
Area of Science:
- Neuroscience
- Immunology
- Pharmacology
Background:
- Peripheral neuronal hypersensitivity, often seen in pain conditions, is linked to interactions between immune cells and neurons.
- Existing in vitro models lack the physiological and anatomical relevance needed to accurately predict human treatment efficacy.
- Dorsal root ganglia (DRG) are crucial in transmitting pain signals, making them a key focus for pain research.
Purpose of the Study:
- To develop and validate a novel, physiologically and anatomically relevant in vitro model for studying dorsal root ganglia (DRG) and neural-immune cell interactions.
- To provide a more accurate platform for investigating pain mechanisms and screening potential pain therapeutics.
- To overcome the limitations of current in vitro models in predicting human treatment outcomes.
Main Methods:
- Harvesting primary DRGs from adult Sprague Dawley rats.
- Culturing intact DRGs within a natural hydrogel in a specialized 48-well, three-compartment plate.
- Isolating DRG cell bodies from their neurites to study specific cellular interactions.
Main Results:
- The multi-compartment system successfully cultured intact DRGs, enabling robust growth.
- The model maintains anatomical relevance by separating cell bodies from neurites and physiological relevance through cell types and mechanical properties.
- This platform facilitates the study of interactions between neural and immune cells in a controlled environment.
Conclusions:
- The developed multi-compartment DRG culture system is a valuable tool for preclinical pain research.
- This advanced in vitro model improves the study of neural-immune interactions relevant to pain.
- The platform offers a more predictive screening approach for pain treatments, potentially improving translation to human efficacy.
