Advances in Human Stem Cell-Derived Neuronal Cell Culturing and Analysis

  • Laura Ylä-Outinen
  • Jarno M. A. Tanskanen
  • Fikret E. Kapucu
  • Anu Hyysalo
  • Jari A. K. Hyttinen
  • Susanna NarkilahtiEmail author
Part of the Advances in Neurobiology book series (NEUROBIOL, volume 22)


This chapter provides an overview of the current stage of human in vitro functional neuronal cultures, their biological application areas, and modalities to analyze their behavior. During the last 10 years, this research area has changed from being practically non-existent to one that is facing high expectations. Here, we present a case study as a comprehensive short history of this process based on extensive studies conducted at NeuroGroup (University of Tampere) and Computational Biophysics and Imaging Group (Tampere University of Technology), ranging from the differentiation and culturing of human pluripotent stem cell (hPSC)-derived neuronal networks to their electrophysiological analysis. After an introduction to neuronal differentiation in hPSCs, we review our work on their functionality and approaches for extending cultures from 2D to 3D systems. Thereafter, we discuss our target applications in neuronal developmental modeling, toxicology, drug screening, and disease modeling. The development of signal analysis methods was required due to the unique functional and developmental properties of hPSC-derived neuronal cells and networks, which separate them from their much-used rodent counterparts. Accordingly, a line of microelectrode array (MEA) signal analysis methods was developed. This work included the development of action potential spike detection methods, entropy-based methods and additional methods for burst detection and quantification, joint analysis of spikes and bursts to analyze the spike waveform compositions of bursts, assessment methods for network synchronization, and computational simulations of synapses and neuronal networks.


Human neurons Human pluripotent stem cells Microelectrode arrays Signal analysis 



The authors would like to thank Tiina Joki, PhD, and Outi Paloheimo, MSc, for their help with figure preparation. This work was funded by the Human Spare Parts Project funded by Business Finland (formerly the Finnish Funding Agency for Technology and Innovation (TEKES)), 3DNeuroN project in the European Union’s Seventh Framework Programme, Future and Emerging Technologies (grant agreement number 296590), Academy of Finland grants (LY grant number 286990 and SN grant numbers 311017 and 312414), and Jane and Aatos Erkko Foundation (JMAT grant “Biological Neuronal Communications and Computing with ICT”).


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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Laura Ylä-Outinen
    • 1
  • Jarno M. A. Tanskanen
    • 2
  • Fikret E. Kapucu
    • 3
    • 4
  • Anu Hyysalo
    • 1
  • Jari A. K. Hyttinen
    • 2
  • Susanna Narkilahti
    • 1
    Email author
  1. 1.NeuroGroup, Faculty of Medicine and Health TechnologyTampere UniversityTampereFinland
  2. 2.Computational Biophysics and Imaging Group, Faculty of Medicine and Health TechnologyTampere UniversityTampereFinland
  3. 3.Danish Research Institute of Translational Neuroscience - DANDRITE, Aarhus UniversityAarhusDenmark
  4. 4.Department of BiomedicineAarhus UniversityAarhusDenmark

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