A Novel Protocol to Derive Cervical Motor Neurons from Induced Pluripotent Stem Cells for Amyotrophic Lateral Sclerosis

Overview

Journal Stem Cell Reports

Publisher Cell Press

Specialty Cell Biology

Date 2023 Aug 18

PMID 37595581

Authors

Meimei Yang

Min Liu

Yajaira Feller Sanchez

Sahar Avazzadeh

Leo R Quinlan

Gang Liu

Yin Lu

Guangming Yang

Timothy OBrien

David C Henshall

Orla Hardiman

Sanbing Shen

Affiliations

Soon will be listed here.

Abstract

Sporadic amyotrophic lateral sclerosis (sALS) is the majority of ALS, and the lack of appropriate disease models has hindered its research. Induced pluripotent stem cell (iPSC) technology now permits derivation of iPSCs from somatic cells of sALS patients to investigate disease phenotypes and mechanisms. Most existing differentiation protocols are time-consuming or low efficient in generating motor neurons (MNs). Here we report a rapid and simple protocol to differentiate MNs in monolayer culture using small molecules, which led to nearly pure neural stem cells in 6 days, robust OLIG2 pMNs (73%-91%) in 12 days, enriched CHAT cervical spinal MNs (sMNs) (88%-97%) in 18 days, and functionally mature sMNs in 28 days. This simple and reproducible protocol permitted the identification of hyperexcitability phenotypes in our sALS iPSC-derived sMNs, and its application in neurodegenerative diseases should facilitate in vitro disease modeling, drug screening, and the development of cell therapy.

Citing Articles

Stem Cell Therapy for the Treatment of Amyotrophic Lateral Sclerosis: Comparison of the Efficacy of Mesenchymal Stem Cells, Neural Stem Cells, and Induced Pluripotent Stem Cells.

Frawley L, Taylor N, Sivills O, McPhillamy E, To T, Wu Y Biomedicines. 2025; 13(1).

PMID: 39857620 PMC: 11763168. DOI: 10.3390/biomedicines13010035.

n-Butylidenephthalide recovered calcium homeostasis to ameliorate neurodegeneration of motor neurons derived from amyotrophic lateral sclerosis iPSCs.

Deng Y, Liu J, Ting H, Kuo T, Chiang C, Lin E PLoS One. 2024; 19(11):e0311573.

PMID: 39509425 PMC: 11542850. DOI: 10.1371/journal.pone.0311573.

Polyethyleneimine facilitates the growth and electrophysiological characterization of iPSC-derived motor neurons.

Yang M, You D, Liu G, Lu Y, Yang G, OBrien T Sci Rep. 2024; 14(1):26106.

PMID: 39478194 PMC: 11525838. DOI: 10.1038/s41598-024-77710-1.

How is Excitotoxicity Being Modelled in iPSC-Derived Neurons?.

Cheng J, Cook A, Talbot J, Perry S Neurotox Res. 2024; 42(5):43.

PMID: 39405005 PMC: 11480214. DOI: 10.1007/s12640-024-00721-3.

A comprehensive review of electrophysiological techniques in amyotrophic lateral sclerosis research.

Ren K, Wang Q, Jiang D, Liu E, Alsmaan J, Jiang R Front Cell Neurosci. 2024; 18:1435619.

PMID: 39280794 PMC: 11393746. DOI: 10.3389/fncel.2024.1435619.

References

Yang M, Liu M, Ding Y, Vajda A, Ma J, Cui H . Generation of twelve induced pluripotent stem cell lines from two healthy controls and two patients with sporadic amyotrophic lateral sclerosis. Stem Cell Res. 2020; 44:101752. DOI: 10.1016/j.scr.2020.101752. View

Wichterle H, Lieberam I, Porter J, Jessell T . Directed differentiation of embryonic stem cells into motor neurons. Cell. 2002; 110(3):385-97. DOI: 10.1016/s0092-8674(02)00835-8. View

Niedermeyer S, Murn M, Choi P . Respiratory Failure in Amyotrophic Lateral Sclerosis. Chest. 2018; 155(2):401-408. DOI: 10.1016/j.chest.2018.06.035. View

Peljto M, Wichterle H . Programming embryonic stem cells to neuronal subtypes. Curr Opin Neurobiol. 2010; 21(1):43-51. PMC: 3050008. DOI: 10.1016/j.conb.2010.09.012. View

Sagner A, Gaber Z, Delile J, Kong J, Rousso D, Pearson C . Olig2 and Hes regulatory dynamics during motor neuron differentiation revealed by single cell transcriptomics. PLoS Biol. 2018; 16(2):e2003127. PMC: 5811045. DOI: 10.1371/journal.pbio.2003127. View

Roelink H, Porter J, Chiang C, Tanabe Y, Chang D, Beachy P . Floor plate and motor neuron induction by different concentrations of the amino-terminal cleavage product of sonic hedgehog autoproteolysis. Cell. 1995; 81(3):445-55. DOI: 10.1016/0092-8674(95)90397-6. View

Chambers S, Fasano C, Papapetrou E, Tomishima M, Sadelain M, Studer L . Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol. 2009; 27(3):275-80. PMC: 2756723. DOI: 10.1038/nbt.1529. View

Avazzadeh S, Quinlan L, Reilly J, McDonagh K, Jalali A, Wang Y . NRXN1α is associated with increased excitability in ASD iPSC-derived neurons. BMC Neurosci. 2021; 22(1):56. PMC: 8442436. DOI: 10.1186/s12868-021-00661-0. View

Avazzadeh S, McDonagh K, Reilly J, Wang Y, Boomkamp S, McInerney V . Increased Ca signaling in neurons derived from ASD induced pluripotent stem cells. Mol Autism. 2020; 10:52. PMC: 6937972. DOI: 10.1186/s13229-019-0303-3. View

10.

Peljto M, Dasen J, Mazzoni E, Jessell T, Wichterle H . Functional diversity of ESC-derived motor neuron subtypes revealed through intraspinal transplantation. Cell Stem Cell. 2010; 7(3):355-66. PMC: 2933095. DOI: 10.1016/j.stem.2010.07.013. View

11.

Qu Q, Li D, Louis K, Li X, Yang H, Sun Q . High-efficiency motor neuron differentiation from human pluripotent stem cells and the function of Islet-1. Nat Commun. 2014; 5:3449. DOI: 10.1038/ncomms4449. View

12.

Pieri M, Albo F, Gaetti C, Spalloni A, Bengtson C, Longone P . Altered excitability of motor neurons in a transgenic mouse model of familial amyotrophic lateral sclerosis. Neurosci Lett. 2003; 351(3):153-6. DOI: 10.1016/j.neulet.2003.07.010. View

13.

Kalani M, Cheshier S, Cord B, Bababeygy S, Vogel H, Weissman I . Wnt-mediated self-renewal of neural stem/progenitor cells. Proc Natl Acad Sci U S A. 2008; 105(44):16970-5. PMC: 2575225. DOI: 10.1073/pnas.0808616105. View

14.

Andrews M, Kong J, Novitch B, Butler S . New perspectives on the mechanisms establishing the dorsal-ventral axis of the spinal cord. Curr Top Dev Biol. 2019; 132:417-450. PMC: 8418640. DOI: 10.1016/bs.ctdb.2018.12.010. View

15.

Patani R . Generating Diverse Spinal Motor Neuron Subtypes from Human Pluripotent Stem Cells. Stem Cells Int. 2016; 2016:1036974. PMC: 4707335. DOI: 10.1155/2016/1036974. View

16.

Lu Q, Sun T, Zhu Z, Ma N, Garcia M, Stiles C . Common developmental requirement for Olig function indicates a motor neuron/oligodendrocyte connection. Cell. 2002; 109(1):75-86. DOI: 10.1016/s0092-8674(02)00678-5. View

17.

Vucic S, Nicholson G, Kiernan M . Cortical hyperexcitability may precede the onset of familial amyotrophic lateral sclerosis. Brain. 2008; 131(Pt 6):1540-50. DOI: 10.1093/brain/awn071. View

18.

van Zundert B, Peuscher M, Hynynen M, Chen A, Neve R, Brown Jr R . Neonatal neuronal circuitry shows hyperexcitable disturbance in a mouse model of the adult-onset neurodegenerative disease amyotrophic lateral sclerosis. J Neurosci. 2008; 28(43):10864-74. PMC: 3844745. DOI: 10.1523/JNEUROSCI.1340-08.2008. View

19.

Portaro S, Morini E, Santoro M, Accorinti M, Marzullo P, Naro A . Breathlessness in amyotrophic lateral sclerosis: A case report on the role of osteoporosis in the worsening of respiratory failure. Medicine (Baltimore). 2018; 97(45):e13026. PMC: 6250541. DOI: 10.1097/MD.0000000000013026. View

20.

Cohen Y, Anaby D, Morozov D . Diffusion MRI of the spinal cord: from structural studies to pathology. NMR Biomed. 2016; 30(3). DOI: 10.1002/nbm.3592. View