Journal: Scientific Reports
Article Title: Bioengineered model of the human motor unit with physiologically functional neuromuscular junctions
doi: 10.1038/s41598-021-91203-5
Figure Lengend Snippet: Number of iPSC motor neuron spheroids loaded in tissue engineered skeletal muscle determines functionality of neuromuscular tissues. ( a ) Schematic detailing method of iPSC motor neuron loading to generate neuromuscular tissues at different stages of myogenic development. (i) MN progenitors are loaded to spheroid microplates and matured for 14 days. (ii) Primary HDMCs are seeded in established type I collagen matrix, loaded into 3D printed mould and cultured to maturity for 2 weeks. Matured MNs are then added within optimised type I collagen hydrogel, seeded around SkM and cultured for a further 2 weeks. MN; Motor neuron, SkM; Skeletal muscle, HDMC; Human derived muscle cells, GM; Growth medium, DM; Differentiation medium, ECM; Extracellular matrix, MM; Motor neuron maintenance medium. ( b ) Addition of iPSC motor neurons with type I collagen perimysium at loading densities of 2, 4 or 6 spheroids per tissue evidence tetanic force and ( c ) concentration dependent spontaneous twitch profiles compared to skeletal muscle only tissues. ( d ) Quantification of functional tetanic and ( e ) spontaneous twitch force output with 2, 4 or 6 motor neuron spheroids. Beginning of representative tetanus and twitch contractions represents initiation of electrical field stimulation, time elapsed before this point is un-stimulated baseline recording. Individual functional data points indicative of n = 3 contraction profiles, totalling minimum of n = 9 and maximum of n = 15 per condition and presented ± standard deviation (SD). **P ≤ 0.01 and ***P ≤ 0.001.
Article Snippet: At cellular confluence, motor neuron progenitors were dissociated using TrypLE select (1x, Gibco, Fisher), centrifuged at 200 G for 5 min and seeded in 96-well U bottom spheroid microplates (MoBiTec, MS-9096UZ) at 1 × 10 4 cells/well or monolayer at 1 × 10 5 cells/cm 2 in motor neuron maintenance medium (MM, Axol) supplemented with 0.5 µM RA, 5 ng/mL brain derived neurotrophic factor (BDNF, PeproTech) and 10 ng/mL ciliary neurotrophic factor (CNTF, Axol).
Techniques: Cell Culture, Derivative Assay, Concentration Assay, Functional Assay, Standard Deviation
Axol Bioscience is a verified supplier 
Axol Bioscience manufactures this product 
