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Bioprinting of Adult Dorsal Root Ganglion (DRG) Neurons Using Laser-Induced Side Transfer (LIST)

Authors: Roversi KEbrahimi Orimi HFalchetti MLummertz da Rocha ETalbot SBoutopoulos C


Affiliations

1 Département de Pharmacologie et Physiologie, Université de Montréal, Montréal, QC H3C 3J7, Canada.
2 Centre de Recherche Hôpital Maisonneuve-Rosemont, Montréal, QC H1T 2M4, Canada.
3 Department of Mechanical, Industrial and Aerospace Engineering, Concordia University, Montréal, QC H3G 1M8, Canada.
4 Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florianópolis 88040-900, Brazil.
5 Département d'Ophtalmologie, Université de Montréal, Montréal, QC H3C 3J7, Canada.
6 Institut de Génie Biomédical, Université de Montréal, Montréal, QC H3C 3J7, Canada.

Description

Cell bioprinting technologies aim to fabricate tissuelike constructs by delivering biomaterials layer-by-layer. Bioprinted constructs can reduce the use of animals in drug development and hold promise for addressing the shortage of organs for transplants. Here, we sought to validate the feasibility of bioprinting primary adult sensory neurons using a newly developed laser-assisted cell bioprinting technology, known as Laser-Induced Side Transfer (LIST). We used dorsal root ganglion neurons (DRG; cell bodies of somatosensory neurons) to prepare our bioink. DRG-laden- droplets were printed on fibrin-coated coverslips and their viability, calcium kinetics, neuropeptides release, and neurite outgrowth were measured. The transcriptome of the neurons was sequenced. We found that LIST-printed neurons maintain high viability (Printed: 86%, Control: 87% on average) and their capacity to release neuropeptides (Printed CGRP: 130 pg/mL, Control CGRP: 146 pg/mL). In addition, LIST-printed neurons do not show differences in the expressed genes compared to control neurons. However, in printed neurons, we found compromised neurite outgrowth and lower sensitivity to the ligand of the TRPV1 channel, capsaicin. In conclusion, LIST-printed neurons maintain high viability and marginal functionality losses. Overall, this work paves the way for bioprinting functional 2D neuron assays.


Keywords: adult DRG neuronscalcium kineticslaser-assisted bioprintinglaser-induced side transfersensory neuronstranscriptomeviability


Links

PubMed: https://pubmed.ncbi.nlm.nih.gov/34442487/

DOI: 10.3390/mi12080865