Amyotrophic Lateral Sclerosis Cell Models

Spinal Motor Neuron Differentiation Protocols

Optimized differentiation protocols are a prerequisite for reliable and predictive disease models. We use spinal motor neurons from BrainXell Inc. with excellent quality parameters. These cells demonstrate an extremely high percentage of spinal motor neurons.

Cultivation and differentiation protocols were optimized using our comprehensive quality control approach. Batches are quality-controlled and perform an internal release.

Electrical activity is a crucial parameter that describes the functional status of a cell culture.

As ALS is a neurodegenerative disease correlated with the age of patients, long-term experiments are essential to produce disease phenotypes; it can be supposed that in vitro models of ALS need a minimum of time to start with pathological mechanisms.

Image: Live-Dead Assay of C9orf72 spinal motor neurons after 14 div, green: living cells, red: dead cells.

Characterization of Cell lines

We continuously characterize our cell lines. We offer after-request pieces of information on the transcriptomics profiles of these cells. Further molecular biomarker characterization can be found here: Molecular readouts or imaging readouts.

Human iPSC-derived Spinal Motor Neurons Mutations

Mutations of C9orf72, SOD1, TDP43, FUS, and others cause familial ALS, which are essential and valuable for ALS disease modeling for phenotypic screening.

The mutations C9orf72, SOD1, TDP43, and FUS cover different pathological mechanisms of ALS, so phenotypic assays with these other mutated iPSC-derived motor neurons will gain predictive power.

C9orf72

The C9orf72 gene is implicated in ALS by repeating GGGGCC expansion in non-coding regions with effects on RNA binding and autophagy regulation. The C9orf72 mutation is also causing, in some cases, frontal temporal dementia.

Mechanisms: RNA processing; nucleocytoplasmic transport defects; proteasome impairment; autophagy; inflammation; protein aggregation; dipeptide repeat proteins (DPRs).

SOD1

The mutations of SOD1 affect the enzyme Cu/Zn-superoxide dismutase 1. These mutations cause the degeneration of motor neurons by oxidative stress, protein aggregates, mitochondrial defects, and glutamate excitotoxicity.

Mechanisms: Dismutates superoxide free radicals; oxidative stress; protein aggregation; mitochondrial dysfunction; axonal transport defects; proteasome impairment; glial dysfunction.

TDP-43

The TAR DNA-binding protein 43 (TDP-43) is another mutation that causes fALS.

Mechanisms: RNA processing; nucleocytoplasmic transport defects; stress granule function; protein aggregation

FUS

Fused in sarcoma RNA binding protein (a component of the hnRNP complex)

Mechanisms: RNA processing; DNA damage repair defects; nucleocytoplasmic transport defects; stress granule function; protein aggregation

ALS iPSC derived Cell-Lines

NeuroProof offers tests with the following ALS cell lines:

Protein	Mutation/Variant	Motor neurons	Spinal astrocytes	Microglia
C9orf72	ND50001	studied by NPS	studied by NPS	on request
SOD1	A4V
	D90A/D90D	studied by NPS	studied by NPS	available/prev. avail.
	H44R/H44H
TDP-43	G298S/G298G	studied by NPS	available/prev. avail.	available/prev. avail.
	A382T/A382A
FUS	H517Q/H517H	available/prev. avail.	in production	in production

Co-Culture Models

NeuroProof offers ALS models as mono-culture, co-culture, or tri-culture from one genotype.

Glial cells are involved in ALS pathology. Co-culture of spinal motor neurons and glial cells are efficient tools for phenotypic modeling of ALS disease in a dish, as is demonstrated in the following example:

Example of a co-culture of C9orf72 spinal motor neurons and C9orf72 astrocytes:

Image: Live-Dead Assay of C9orf72 spinal motor neurons after 8 div, green: living cells, red: dead cells.

Image: Live-Dead Assay of C9orf72 astrocytes after 8 div, green: living cells, red: dead cells.

Image: Live-Dead Assay of a co-culture of C9orf72 spinal motor neurons and astrocytes after 8 div, green living cells, red dead cells.

Tailored ALS Cell Models

Select your specific ALS cell model from different mutations, co-cultures, and timing protocols or combine them with an additional chemical challenge such as beta-methylamino-L-alanine (BMAA).

Get in Touch

NeuroProof is continuously expanding its assay portfolio, asking us for potential new ALS cell lines with familial or sporadic forms and co-cultures with astrocytes and microglia. NeuroProof performs integrated projects, from sourcing diseased cell lines, cell differentiation, and assay development to compound screening campaigns..