Stromal cell-derived inducing activity, Nurr1, and signaling molecules synergistically induce dopaminergic neurons from mouse embryonic stem cells

被引:65
作者
Kim, Dong-Wook
Chung, Sangmi
Hwang, Mikyeong
Ferree, Andrew
Tsai, Hsing-Chen
Park, Jae-Joon
Chung, Seungsoo
Nam, Taick Sang
Kang, Un Jung
Isacson, Ole
Kim, Kwang-Soo
机构
[1] Harvard Univ, McLean Hosp, Sch Med, Udall Parkinsons Dis Res Ctr Excellence, Belmont, MA 02178 USA
[2] Harvard Univ, McLean Hosp, Sch Med, Mol Neurobiol Labs, Belmont, MA 02178 USA
[3] Yonsei Univ, Coll Med, Dept Physiol, Seoul 120749, South Korea
[4] Harvard Univ, McLean Hosp, Sch Med, Neuroregenerat Labs, Belmont, MA 02178 USA
[5] Univ Chicago, Dept Neurol & Pharmacol, Chicago, IL 60637 USA
[6] Univ Chicago, Dept Physiol, Chicago, IL 60637 USA
[7] Univ Chicago, Dept Neurobiol, Chicago, IL 60637 USA
关键词
embryonic stem cells; stromal cell-derived inducing activity; Nurr1; dopaminergic neurons; Parkinson's disease;
D O I
10.1634/stemcells.2005-0233
中图分类号
Q813 [细胞工程];
学科分类号
摘要
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 (similar to twofold) than the naive 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 (similar to 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.
引用
收藏
页码:557 / 567
页数:11
相关论文
共 46 条
  • [41] Fate of mesencephalic AHD2-expressing dopamine progenitor cells in Nurr1 mutant mice
    Wallén, Å
    Zetterström, RH
    Solomin, L
    Arvidsson, M
    Olson, L
    Perlmann, T
    [J]. EXPERIMENTAL CELL RESEARCH, 1999, 253 (02) : 737 - 746
  • [42] Directed differentiation of embryonic stem cells into motor neurons
    Wichterle, H
    Lieberam, I
    Porter, JA
    Jessell, TM
    [J]. CELL, 2002, 110 (03) : 385 - 397
  • [43] FGF and Shh signals control dopaminergic and serotonergic cell fate in the anterior neural plate
    Ye, WL
    Shimamura, K
    Rubenstein, JLR
    Hynes, MA
    Rosenthal, A
    [J]. CELL, 1998, 93 (05) : 755 - 766
  • [44] Dopaminergic differentiation of human embryonic stem cells
    Zeng, XM
    Cai, JL
    Chen, J
    Luo, YQ
    You, ZB
    Fotter, E
    Wang, Y
    Harvey, B
    Miura, T
    Backman, C
    Chen, GJ
    Rao, MS
    Freed, WJ
    [J]. STEM CELLS, 2004, 22 (06) : 925 - 940
  • [45] Dopamine neuron agenesis in Nurr1-deficient mice
    Zetterstrom, RH
    Solomin, L
    Jansson, L
    Hoffer, BJ
    Olson, L
    Perlmann, T
    [J]. SCIENCE, 1997, 276 (5310) : 248 - 250
  • [46] In vitro differentiation of transplantable neural precursors from human embryonic stem cells
    Zhang, SC
    Wernig, M
    Duncan, ID
    Brüstle, O
    Thomson, JA
    [J]. NATURE BIOTECHNOLOGY, 2001, 19 (12) : 1129 - 1133