Open AccessStromal Cell–Derived Inducing Activity, Nurr1, and Signaling Molecules Synergistically Induce Dopaminergic Neurons from Mouse Embryonic Stem Cells
Author(s) -
Kim DongWook,
Chung Sangmi,
Hwang Mikyeong,
Ferree Andrew,
Tsai HsingChen,
Park JaeJoon,
Chung Seungsoo,
Nam Taick Sang,
Kang Un Jung,
Isacson Ole,
Kim KwangSoo
Publication year - 2006
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2005-0233
Subject(s) - biology , embryonic stem cell , microbiology and biotechnology , dopaminergic , transplantation , stromal cell , tyrosine hydroxylase , cellular differentiation , dopamine transporter , dopamine , stem cell , ascorbic acid , cell type , medicine , cell , endocrinology , cancer research , biochemistry , gene , food science
To induce differentiation of embryonic stem cells (ESCs) into specialized cell types for therapeutic purposes, it may be desirable to combine genetic manipulation and appropriate differentiation signals. We studied the induction of dopaminergic (DA) neurons from mouse ESCs by overexpressing the transcription factor Nurr1 and coculturing with PA6 stromal cells. Nurr1‐expressing ESCs (N2 and N5) differentiated into a higher number of neurons (∼twofold) than the naïve ESCs (D3). In addition, N2/N5‐derived cells contained a significantly higher proportion (>50%) of tyrosine hydroxylase (TH) + neurons than D3 (<30%) and an even greater proportion of TH + neurons (∼90%) when treated with the signaling molecules sonic hedgehog, fibroblast growth factor 8, and ascorbic acid. N2/N5‐derived cells express much higher levels of DA markers (e.g., TH, dopamine transporter, aromatic amino acid decarboxylase, and G protein–regulated inwardly rectifying K + channel 2) and produce and release a higher level of dopamine, compared with D3‐derived cells. Furthermore, the majority of generated neurons exhibited electrophysiological properties characteristic of midbrain DA neurons. Finally, transplantation experiments showed efficient in vivo integration/generation of TH + neurons after implantation into mouse striatum. Taken together, our results show that the combination of genetic manipulation(s) and in vitro cell differentiation conditions offers a reliable and effective induction of DA neurons from ESCs and may pave the way for future cell transplantation therapy in Parkinson's disease.