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Article

Generation of Active Neurons from Mouse Embryonic Stem Cells Using Retinoic Acid and Purmorphamine

by
Ruby Vajaria
1,
DeAsia Davis
1,
Francesco Tamagnini
2,
Duncan G. G. McMillan
1,
Nandini Vasudevan
1 and
Evangelos Delivopoulos
1,*
1
School of Biological Sciences, University of Reading, Reading RG6 6UB, UK
2
School of Pharmacy, University of Reading, Reading RG6 6UR, UK
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2025, 26(17), 8372; https://doi.org/10.3390/ijms26178372 (registering DOI)
Submission received: 19 June 2025 / Revised: 14 August 2025 / Accepted: 25 August 2025 / Published: 28 August 2025
(This article belongs to the Special Issue Neural Stem Cells: Latest Applications and Future Perspectives)

Abstract

Multiple differentiation protocols have emerged in recent years, producing neurons with diverse morphologies, gene and protein expression profiles, and functionality. Many of these differentiation techniques require months of culture and the use of expensive growth factors. Most importantly, the derived neurons usually do not exhibit any electrical activity. This limits the value of the protocol as a tool for engineering and investigating neural networks. Here, we describe an efficacious method for differentiating mouse embryonic stem cells into functional neurons. CGR8 cells were neurally induced via the simultaneous application of retinoic acid and purmorphamine. The derived cells expressed neuronal (TUJ1 and NeuN) and synaptic (GAD2, PSD-95, Synaptophysin, and VGLUT1) markers. During whole-cell recordings, neurons exhibited inward and outward currents, likely caused by fast-inactivating voltage-gated potassium channels. Upon current injection, miniature action potentials were also recorded. The efficient generation of diverse subtypes of functional neurons can be a useful tool in fundamental investigations of neural network activity and translational studies.
Keywords: embryonic stem cells (ESCs); neural differentiation; leukemia inhibitory factor (LIF); retinoic acid; cell culture; neuron embryonic stem cells (ESCs); neural differentiation; leukemia inhibitory factor (LIF); retinoic acid; cell culture; neuron

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MDPI and ACS Style

Vajaria, R.; Davis, D.; Tamagnini, F.; McMillan, D.G.G.; Vasudevan, N.; Delivopoulos, E. Generation of Active Neurons from Mouse Embryonic Stem Cells Using Retinoic Acid and Purmorphamine. Int. J. Mol. Sci. 2025, 26, 8372. https://doi.org/10.3390/ijms26178372

AMA Style

Vajaria R, Davis D, Tamagnini F, McMillan DGG, Vasudevan N, Delivopoulos E. Generation of Active Neurons from Mouse Embryonic Stem Cells Using Retinoic Acid and Purmorphamine. International Journal of Molecular Sciences. 2025; 26(17):8372. https://doi.org/10.3390/ijms26178372

Chicago/Turabian Style

Vajaria, Ruby, DeAsia Davis, Francesco Tamagnini, Duncan G. G. McMillan, Nandini Vasudevan, and Evangelos Delivopoulos. 2025. "Generation of Active Neurons from Mouse Embryonic Stem Cells Using Retinoic Acid and Purmorphamine" International Journal of Molecular Sciences 26, no. 17: 8372. https://doi.org/10.3390/ijms26178372

APA Style

Vajaria, R., Davis, D., Tamagnini, F., McMillan, D. G. G., Vasudevan, N., & Delivopoulos, E. (2025). Generation of Active Neurons from Mouse Embryonic Stem Cells Using Retinoic Acid and Purmorphamine. International Journal of Molecular Sciences, 26(17), 8372. https://doi.org/10.3390/ijms26178372

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