Research Article

Similarly derived and cultured hESC lines show variation in their developmental potential towards neuronal cells in long-term culture

Regea Institute for Regenerative Medicine, University of Tampere & Tampere University Hospital, Tampere, Finland; Institute for Regenerative Medicine, University of Tampere, Biokatu 12, 6th floor, 33520 Tampere, Finland

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Minna Salomäki

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Laura Ylä-Outinen

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Teemu J Heikkilä

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Jari AK Hyttinen

Department of Biomedical Engineering, Tampere University of Technology, Tampere, Finland

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Harri Pihlajamäki

Centre for Military Medicine, Helsinki, Finland

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Riitta Suuronen

Department of Biomedical Engineering, Tampere University of Technology, Tampere, Finland

Department of Eye, Ear & Oral Diseases, Tampere University Hospital, Tampere, Finland

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Heli Skottman

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Susanna Narkilahti

† Author for correspondence

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Email the corresponding author at susanna.narkilahti@regea.fi

Published Online:27 Sep 2010https://doi.org/10.2217/rme.10.58

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Abstract

Background: Human embryonic stem cells (hESCs) can differentiate into any human cell type, including CNS cells, and thus have high potential in regenerative medicine. Several protocols exist for neuronal differentiation of hESCs, which do not necessarily work for all hESC lines. Materials & methods: We tested the differentiation capacity of four similarly derived and cultured hESC lines (HS181, HS360, HS362 and HS401) in suspension culture in relatively simple neural differentiation medium for up to 20 weeks. Results: All the hESC lines differentiated into neuronal cells, but in a line-dependent manner. Using our method, the HS181- and HS360-derived neurospheres differentiated in vitro into pure neuronal cell populations within 6 weeks, whereas HS362 and HS401 reached their peak of differentiation in 12 weeks, but never produced pure neuronal cell populations using the present method. The withdrawal of FGF from suspension culture increased the in vitro differentiation potential. The hESC-derived neurospheres formed functional neuronal networks when replated on a microelectrode array and responded as expected to pharmacologic modulation. Conclusion: Simple neurosphere culture is a suitable method for producing hESC-derived neuronal cells that can form functional neuronal networks from a number of hESC lines. The variation in the differentiation potential of hESC lines into neuronal cells must be carefully considered by those comparing various differentiation methods and designing transplantation therapies for neuronal disorders.

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Published online 27 September 2010

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Acknowledgements

We thank Prof O Hovatta (Karolinska Institute, Sweden) for providing the hESC lines. We thank the personnel of Regea – Institute for Regenerative Medicine for their support and assistance in the stem cell research. University statistician H Huhtala is acknowledged for advice in statistical analyses.

Financial & competing interests disclosure

This study was supported by the Academy of Finland, The Competitive Research Fund of the Pirkanmaa Hospital District, the Finnish Cultural foundation, Maire Taponen Foundation, Orion-Farmos Foundation, the City of Tampere and the University of Tampere. MEA system was funded by BioneXt Tampere. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Writing assistance was utilized in the production of this manuscript. The manuscript was checked by SciTechEdit.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

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