- 作者: Mingming Li, Yu Zou, Qiqi Lu, Ning Tang, Alexis Heng, Intekhab Islam, Huei Jinn Tong, Gavin S. Dawe and Tong Cao
- 作者服務機構: Faculty of Dentistry, National University of Singapore, Kent Ridge, Singapore
- 中文摘要: --
- 英文摘要:
Background
Parkinson’s disease (PD) is a severe neurodegenerative disease associated with loss of dopaminergic neurons. Derivation of dopaminergic neurons from human embryonic stem cells (hESCs) could provide new therapeutic options for PD therapy. Dopaminergic neurons are derived from SOX− floor plate (FP) cells during embryonic development in many species and in human cell culture in vitro. Early treatment with sonic hedgehog (Shh) has been reported to efficiently convert hESCs into FP lineages.
Methods
In this study, we attempted to utilize a Shh-free approach in deriving SOX1− FP cells from hESCs in vitro. Neuroectoderm conversion from hESCs was achieved with dual inhibition of the BMP4 (LDN193189) and TGF-β signaling pathways (SB431542) for 24 h under defined culture conditions.
Results
Following a further 5 days of treatment with LDN193189 or LDN193189 + SB431542, SOX1− FP cells constituted 70–80 % of the entire cell population. Upon treatment with Shh and FGF8, the SOX1− FP cells were efficiently converted to functional Nurr1+ and TH+ dopaminergic cells (patterning), which constituted more than 98 % of the entire cell population. However, when the same growth factors were applied to SOX1+ cells, only less than 4 % of the cells became Nurr1+, indicating that patterning was effective only if SOX1 expression was down-regulated. After transplanting the Nurr1+ and TH+ cells into a hemiparkinsonian rat model, significant improvements were observed in amphetamine induced ipslateral rotations, apomorphine induced contra-lateral rotations and Rota rod motor tests over a duration of 8 weeks.
Conclusions
Our findings thus provide a convenient approach to FP development and functional dopaminergic neuron derivation. - 中文關鍵字: --
- 英文關鍵字: Human embryonic stem cells, Neural conversion, Floor plate, LDN193189, SB431541, PD173074, Dopaminergic neurons