Comparative study of efficacy of dopaminergic neuron differentiation between embryonic stem cell and protein-based induced pluripotent stem cell

Yoo-Wook Kwon; Yeon-Ju Chung; Joonoh Kim; Ho-Jae Lee; Jihwan Park; Tae-Young Roh; Hyun-Jai Cho; Chang-Hwan Yoon; Bon-Kwon Koo; Hyo-Soo Kim

doi:10.1371/journal.pone.0085736

Comparative study of efficacy of dopaminergic neuron differentiation between embryonic stem cell and protein-based induced pluripotent stem cell

PLoS One. 2014 Jan 22;9(1):e85736. doi: 10.1371/journal.pone.0085736. eCollection 2014.

Authors

Affiliations

¹ National Research Laboratory for Stem Cell Niche, Seoul National University Hospital, Seoul, Korea ; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea.
² Division of Molecular and Life Sciences,Pohang University of Science and Technology, Pohang, Korea.
³ National Research Laboratory for Stem Cell Niche, Seoul National University Hospital, Seoul, Korea ; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea ; Department of Internal Medicine, Seoul National University, Seoul, Korea.
⁴ Cardiovascular center, Seoul National University Bundang Hospital, Seoul National University, Seoul, Korea.
⁵ Department of Internal Medicine, Seoul National University, Seoul, Korea.
⁶ National Research Laboratory for Stem Cell Niche, Seoul National University Hospital, Seoul, Korea ; Innovative Research Institute for Cell Therapy, Seoul National University Hospital, Seoul, Korea ; Department of Internal Medicine, Seoul National University, Seoul, Korea ; Molecular Medicine and Biopharmaceutical Sciences, Seoul National University, Seoul, Korea.

Abstract

In patients with Parkinson's disease (PD), stem cells can serve as therapeutic agents to restore or regenerate injured nervous system. Here, we differentiated two types of stem cells; mouse embryonic stem cells (mESCs) and protein-based iPS cells (P-iPSCs) generated by non-viral methods, into midbrain dopaminergic (mDA) neurons, and then compared the efficiency of DA neuron differentiation from these two cell types. In the undifferentiated stage, P-iPSCs expressed pluripotency markers as ES cells did, indicating that protein-based reprogramming was stable and authentic. While both stem cell types were differentiated to the terminally-matured mDA neurons, P-iPSCs showed higher DA neuron-specific markers' expression than ES cells. To investigate the mechanism of the superior induction capacity of DA neurons observed in P-iPSCs compared to ES cells, we analyzed histone modifications by genome-wide ChIP sequencing analysis and their corresponding microarray results between two cell types. We found that Wnt signaling was up-regulated, while SFRP1, a counter-acting molecule of Wnt, was more suppressed in P-iPSCs than in mESCs. In PD rat model, transplantation of neural precursor cells derived from both cell types showed improved function. The present study demonstrates that P-iPSCs could be a suitable cell source to provide patient-specific therapy for PD without ethical problems or rejection issues.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation*
Cell Movement
Cells, Cultured
Dopaminergic Neurons / cytology*
Dopaminergic Neurons / metabolism
Dopaminergic Neurons / transplantation
Embryonic Stem Cells / cytology*
Embryonic Stem Cells / metabolism
Female
Gene Expression
Induced Pluripotent Stem Cells / cytology*
Induced Pluripotent Stem Cells / metabolism
Intercellular Signaling Peptides and Proteins / genetics
Membrane Proteins / genetics
Mice
Mice, 129 Strain
Mice, Inbred C57BL
Microscopy, Confocal
Oxidopamine
Parkinson Disease, Secondary / chemically induced
Parkinson Disease, Secondary / surgery
Proteins / metabolism
Rats
Rats, Sprague-Dawley
Reverse Transcriptase Polymerase Chain Reaction
Treatment Outcome
Wnt Proteins / genetics
Wnt Signaling Pathway
Wnt-5a Protein

Substances

Intercellular Signaling Peptides and Proteins
Membrane Proteins
Proteins
Sfrp1 protein, mouse
Wnt Proteins
Wnt-5a Protein
Wnt5a protein, mouse
Oxidopamine

Grants and funding

This study was supported by a grant from the Innovative Research Institute for Cell Therapy (A062260) and the Bio & Medical Technology Development Program of the National Research Foundation funded by the Korean government (MEST) (2010-0020258) and a grant of the Korean Health Technology Research & Development Project, Ministry for Health, Welfare & Family Affairs, Republic of Korea (A100476). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.