Abstract
Several studies have shown that human induced pluripotent stem cell (iPSC)-derivatives are essentially fetal in terms of their maturational status. Inducing ageing in iPSC-motor neuron (MN) models of amyotrophic lateral sclerosis (ALS) has the potential to capture pathology with higher fidelity and consequently improve translational success. We show here that the telomerase inhibitor BIBR1532, hypothesised to recapitulate the telomere attrition hallmark of ageing in iPSC-MNs, was in fact cytotoxic to feeder-free iPSCs when used at doses previously shown to be effective in iPSCs grown on a layer of mouse embryonic fibroblasts. Toxicity in feeder-free cultures was not rescued by co-treatment with Rho Kinase (ROCK) inhibitor (Y-27632). Moreover, the highest concentration of BIBR1532 compatible with continued iPSC culture proved insufficient to induce detectable telomerase inhibition. Our data suggest that direct toxicity by BIBR1532 is the most likely cause of iPSC death observed, and that culture methods may influence enhanced toxicity. Therefore, recapitulation of ageing hallmarks in iPSC-MNs, which might reveal novel and relevant human disease targets in ALS, is not achievable in feeder-free culture through the use of this small molecule telomerase inhibitor.
Highlights
An unequivocal consequence of an ageing human population is an increase in ageassociated neurodegenerative disease prevalence, including amyotrophic lateral sclerosis (ALS)
We found that the treatment of feeder-free induced pluripotent stem cell (iPSC) with BIBR1532 was cytotoxic across a range of concentrations, which was not rescued by ROCK inhibition (Y-27632)
Cultures were treated for 14 days with BIBR1532 at the pluripotent stage and 18 days during differentiation to motor neuron (MN) progenitors, after which treatment was removed for terminal differentiation to
Summary
An unequivocal consequence of an ageing human population is an increase in ageassociated neurodegenerative disease prevalence, including amyotrophic lateral sclerosis (ALS). This is a rapidly progressive degenerative disease of motor neurons (MNs) where patients lose the ability to eat, speak, locomote, and breathe, causing death within 2–5 years of diagnosis. Patient-specific human induced pluripotent stem cells (iPSCs) can differentiate into progeny from any of the three germ layers via developmentally rationalised directed differentiation paradigms [6], including ectodermally-derived MNs. By maintaining patient disease genetics, the iPSC-MN model can capture disease phenotypes in vitro [7,8,9,10]
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