In Vitro Generation of Motor Neuron Precursors From Mouse Embryonic Stem Cells Using Mesoporous Nanoparticles

Abstract

Aim: Stem cell-derived motor neurons (MNs) are utilized to develop replacement strategies for spinal cord disorders. Differentiation of embryonic stem cells into MN precursors involves factors and their repeated administration. We investigated if delivery of factors loaded into mesoporous nanoparticles could be effective for stem cell differentiation in vitro. Materials & methods: We used a mouse embryonic stem cell line expressing green fluorescent protein under the promoter for the MN-specific gene Hb9 to visualize the level of MN differentiation. The differentiation of stem cells was evaluated by expression of MN-specific transcription factors monitored by quantitative real-time PCR reactions and immunocytochemistry. Results: Mesoporous nanoparticles have strong affiliation to the embryoid bodies, penetrate inside the embryoid bodies and come in contact with differentiating cells. Conclusion: Repeated administration of soluble factors into a culture medium can be avoided due to a sustained release effect using mesoporous silica.

Keywords::

• cell transplant
• differentiation
• mesoporous material
• nanomedicine
• stem cell

Supplementary Material

Financial & competing interests disclosure

This work was supported by the Swedish Research Council, projects 2010–20716 (EK), and 2009–4716 (AEGB), Stiftelsen Olle Engkvist Byggare, Signhild Engkvist‘s Stiftelse and the Swedish Institute‘s Visby program Dnr 00613/2011. AE Garcia-Bennett is the cofounder of Nanologica AB (Stockholm, Sweden) a company commercializing nanoporous materials for biomedical and related applications and, therefore, has a potential conflict of interest. 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.

No writing assistance was utilized in the production of this manuscript.

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.

Acknowledgements

We are grateful to Kevin Eggan for generously providing the Hb9::GFP cell line; Zhanna Alekseenko, Elisabet Andersson and Petra Sekyrenko for helping establish embryonic stem cell cultures; Rizwan Qaisar and Stina Höglund for help with confocal imaging; Xin Xia and Natalia Kupferschmidt for assistance with the release measurements of soluble factors; and Ninnie Abrahamsson for technical assistance.

Additional information

Funding

This work was supported by the Swedish Research Council, projects 2010–20716 (EK), and 2009–4716 (AEGB), Stiftelsen Olle Engkvist Byggare, Signhild Engkvist‘s Stiftelse and the Swedish Institute‘s Visby program Dnr 00613/2011. AE Garcia-Bennett is the cofounder of Nanologica AB (Stockholm, Sweden) a company commercializing nanoporous materials for biomedical and related applications and, therefore, has a potential conflict of interest. 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.