Culture of Human iPSC-Derived Motoneurons in Compartmentalized Microfluidic Devices and Quantitative Assays for Studying Axonal Phenotypes

Maria Giovanna Garone; Chiara D'Antoni; Alessandro Rosa

doi:10.1007/978-1-0716-1979-7_12

Culture of Human iPSC-Derived Motoneurons in Compartmentalized Microfluidic Devices and Quantitative Assays for Studying Axonal Phenotypes

Methods Mol Biol. 2022:2429:189-199. doi: 10.1007/978-1-0716-1979-7_12.

Authors

Maria Giovanna Garone¹, Chiara D'Antoni^{1

2}, Alessandro Rosa^{3

4

5}

Affiliations

¹ Department of Biology and Biotechnologies "Charles Darwin", Sapienza University of Rome, Rome, Italy.
² Laboratory Affiliated to Istituto Pasteur Italia-Fondazione Cenci Bolognetti, Department of Biology and Biotechnologies "Charles Darwin", Sapienza University of Rome, Rome, Italy.
³ Department of Biology and Biotechnologies "Charles Darwin", Sapienza University of Rome, Rome, Italy. [email protected].
⁴ Laboratory Affiliated to Istituto Pasteur Italia-Fondazione Cenci Bolognetti, Department of Biology and Biotechnologies "Charles Darwin", Sapienza University of Rome, Rome, Italy. [email protected].
⁵ Center for Life Nano- & Neuro-Science, Fondazione Istituto Italiano di Tecnologia, Rome, Italy. [email protected].

PMID: 35507162
DOI: 10.1007/978-1-0716-1979-7_12

Abstract

In order to use induced Pluripotent Stem Cells (iPSCs) to model neurodegenerative diseases, efficient and homogeneous generation of neurons in vitro represents a key step. Here we describe a method to obtain and characterize functional human spinal and cranial motoneurons using a combined approach of microfluidic chips and programs designed for scientific multidimensional imaging. We have used this approach to analyze axonal phenotypes. These tools are useful to investigate the cellular and molecular bases of neuromuscular diseases, including amyotrophic lateral sclerosis and spinal muscular atrophy.

Keywords: Amyotrophic lateral sclerosis; Axon; Axotomy; Cranial motoneuron; Differentiation protocol; Microfluidic device; Spinal motoneuron; iPSC; piggyBac.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amyotrophic Lateral Sclerosis*
Axons / physiology
Humans
Induced Pluripotent Stem Cells*
Lab-On-A-Chip Devices
Motor Neurons
Phenotype