Next Article in Journal
Markers of Regenerative Processes in Patients with Bipolar Disorder: A Case-control Study
Previous Article in Journal
Evaluating the Impact of Age, Acoustic Exposure, and Electrical Stimulation on Binaural Sensitivity in Adult Bilateral Cochlear Implant Patients
Open AccessArticle

Optimized Protocol to Generate Spinal Motor Neuron Cells from Induced Pluripotent Stem Cells from Charcot Marie Tooth Patients

1
CHU de Limoges, Service de Biochimie et Génétique Moléculaire, F-87000 Limoges, France
2
Université de Limoges, Maintenance Myélinique et Neuropathies Périphériques, EA6309, F-87000 Limoges, France
3
Université de Nantes, RMeS Regenerative Medicine and Skeleton, ONIRIS, INSERM UMR 1229, F-44042 Nantes, France
4
Bogomoletz Institute of Physiology, Department of Sensory Signaling, 01601 Kyiv, Ukraine
5
CHU de Limoges, Service de Neurologie, F-87000 Limoges, France
6
CHU de Limoges, Service de Cytogénétique, F-87000 Limoges, France
7
CHU Henri-Mondor, Département de Génétique, F-94000 Créteil, France
8
Université Paris-Est-Créteil, Inserm U955-E10, F-94000 Créteil, France
9
CHU de Limoges, UF de Bioinformatique, F-87000 Limoges, France
*
Author to whom correspondence should be addressed.
Brain Sci. 2020, 10(7), 407; https://doi.org/10.3390/brainsci10070407
Received: 29 May 2020 / Revised: 18 June 2020 / Accepted: 24 June 2020 / Published: 27 June 2020
(This article belongs to the Section Molecular and Cellular Neuroscience)
Modelling rare neurogenetic diseases to develop new therapeutic strategies is highly challenging. The use of human-induced pluripotent stem cells (hiPSCs) is a powerful approach to obtain specialized cells from patients. For hereditary peripheral neuropathies, such as Charcot–Marie–Tooth disease (CMT) Type II, spinal motor neurons (MNs) are impaired but are very difficult to study. Although several protocols are available to differentiate hiPSCs into neurons, their efficiency is still poor for CMT patients. Thus, our goal was to develop a robust, easy, and reproducible protocol to obtain MNs from CMT patient hiPSCs. The presented protocol generates MNs within 20 days, with a success rate of 80%, using specifically chosen molecules, such as Sonic Hedgehog or retinoic acid. The timing and concentrations of the factors used to induce differentiation are crucial and are given hereby. We then assessed the MNs by optic microscopy, immunocytochemistry (Islet1/2, HB9, Tuj1, and PGP9.5), and electrophysiological recordings. This method of generating MNs from CMT patients in vitro shows promise for the further development of assays to understand the pathological mechanisms of CMT and for drug screening. View Full-Text
Keywords: induced pluripotent stem cells; hiPSC; spinal motor neurons; cellular models; peripheral nervous system; Charcot-Marie-Tooth; CMT; peripheral neuropathy induced pluripotent stem cells; hiPSC; spinal motor neurons; cellular models; peripheral nervous system; Charcot-Marie-Tooth; CMT; peripheral neuropathy
Show Figures

Figure 1

MDPI and ACS Style

Faye, P.-A.; Vedrenne, N.; Miressi, F.; Rassat, M.; Romanenko, S.; Richard, L.; Bourthoumieu, S.; Funalot, B.; Sturtz, F.; Favreau, F.; Lia, A.-S. Optimized Protocol to Generate Spinal Motor Neuron Cells from Induced Pluripotent Stem Cells from Charcot Marie Tooth Patients. Brain Sci. 2020, 10, 407. https://doi.org/10.3390/brainsci10070407

AMA Style

Faye P-A, Vedrenne N, Miressi F, Rassat M, Romanenko S, Richard L, Bourthoumieu S, Funalot B, Sturtz F, Favreau F, Lia A-S. Optimized Protocol to Generate Spinal Motor Neuron Cells from Induced Pluripotent Stem Cells from Charcot Marie Tooth Patients. Brain Sciences. 2020; 10(7):407. https://doi.org/10.3390/brainsci10070407

Chicago/Turabian Style

Faye, Pierre-Antoine; Vedrenne, Nicolas; Miressi, Federica; Rassat, Marion; Romanenko, Sergii; Richard, Laurence; Bourthoumieu, Sylvie; Funalot, Benoît; Sturtz, Franck; Favreau, Frederic; Lia, Anne-Sophie. 2020. "Optimized Protocol to Generate Spinal Motor Neuron Cells from Induced Pluripotent Stem Cells from Charcot Marie Tooth Patients" Brain Sci. 10, no. 7: 407. https://doi.org/10.3390/brainsci10070407

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Search more from Scilit
 
Search
Back to TopTop