A novel 3D bilayer hydrogel tri-culture system for studying functional motor units

Cell & Bioscience

Methodology

Sang-Hyun Oh¹  Dong Jun Lee¹  Jennifer Nhieu²  Thomas Lerdall²  Li-Na Wei²  Chin-Wen Wei²  Liming Milbauer²  Stanley Thayer²  Yu-Lung Lin³ 

[1] Department of Electrical and Computer Engineering, University of Minnesota, 55455, Minneapolis, MN, USA;Department of Pharmacology, University of Minnesota Medical School, 55455, Minneapolis, MN, USA;Department of Pharmacology, University of Minnesota Medical School, 55455, Minneapolis, MN, USA;The Ph.D. Program for Translational Medicine, College of Medical Sciences and Technology, Taipei Medical University, Taipei, Taiwan;International Ph.D. Program for Translational Science, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan;

关键词: Motor unit;    Motor neuron;    Muscle;    Schwann cell;    Neuromuscular;    Hydrogel;    Co-culture;    Motor neuron disorders;   

DOI  :  10.1186/s13578-023-01115-2

 received in 2023-06-07, accepted in 2023-08-27,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundA motor unit (MU) is formed by a single alpha motor neuron (MN) and the muscle fibers it innervates. The MU is essential for all voluntary movements. Functional deficits in the MU result in neuromuscular disorders (NMDs). The pathological mechanisms underlying most NMDs remain poorly understood, in part due to the lack of in vitro models that can comprehensively recapitulate multistage intercellular interactions and physiological function of the MU.ResultsWe have designed a novel three-dimensional (3D) bilayer hydrogel tri-culture system where architecturally organized MUs can form in vitro. A sequential co-culture procedure using the three cell types of a MU, MN, myoblast, and Schwann cell was designed to construct a co-differentiating tri-culture on a bilayer hydrogel matrix. We utilized a µ-molded hydrogel with an additional Matrigel layer to form the bilayer hydrogel device. The µ-molded hydrogel layer provides the topological cues for myoblast differentiation. The Matrigel layer, with embedded Schwann cells, not only separates the MNs from myoblasts but also provides a proper micro-environment for MU development. The completed model shows key MU features including an organized MU structure, myelinated nerves, aligned myotubes innervated on clustered neuromuscular junctions (NMJs), MN-driven myotube contractions, and increases in cytosolic Ca2+ upon stimulation.ConclusionsThis organized and functional in vitro MU model provides an opportunity to study pathological events involved in NMDs and peripheral neuropathies, and can serve as a platform for physiological and pharmacological studies such as modeling and drug screening. Technically, the rational of this 3D bilayer hydrogel co-culture system exploits multiple distinct properties of hydrogels, facilitating effective and efficient co-culturing of diverse cell types for tissue engineering.

【 授权许可】

CC BY   
© Society of Chinese Bioscientists in America (SCBA) 2023

【 图 表 】

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