Generation of Tripotent Neural Progenitor Cells from Rat Embryonic Stem Cells

Zhenkun Wang^{a, b, d, #},
Chao Sheng^{a, b, #},
Tianda Li^{a, b, #},
Fei Teng^{b, c, #},
Lisi Sang^{b, c},
Fenglin Cao^d,
Ziwei Wang^b,
Wanwan Zhu^b,
Wei Li^{b, c},
Xiaoyang Zhao^b,
Zhonghua Liu^a,
Liu Wang^{b
, ,},
Qi Zhou^{b
, ,}

a.
College of Life Science, Northeast Agricultural University of China, Harbin 150030, China
b.
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
c.
Graduate School of Chinese Academy of Sciences, Beijing 100039, China
d.
Central Laboratory, The First Affiliated Hospital, Harbin Medical University, Harbin 150030, China

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Corresponding author: E-mail address: wangliu@ioz.ac.cn (Liu Wang); E-mail address: qzhou@ioz.ac.cn (Qi Zhou)

These authors contributed equally to this work.

摘要

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Abstract: Rat is a valuable model for pharmacological and physiological studies. Germline-competent rat embryonic stem (rES) cell lines have been successfully established and the molecular networks maintaining the self-renewing, undifferentiated state of rES cells have also been well uncovered. However, little is known about the differentiation strategies and the underlying mechanisms of how these authentic rat pluripotent stem cells give rise to specific cell types. The aim of this study is to investigate the neural differentiation capacity of rES cells. By means of a modified procedure based on previous publications – combination of mitogen-activated protein kinase (MAPK) and glycogen synthase kinase 3 (GSK3) inhibitors (two inhibitors, “2i”) with feeder-conditioned medium, we successfully obtained high-quality rat embryoid bodies (rEBs) from rES cells and then differentiated them to tripotent neural progenitors. These rES cell-derived neural progenitor cells (rNPCs) were capable of self-renewing and giving rise to all three neural lineages, including astrocytes, oligodendrocytes, and neurons. Besides, these rES cell-derived neurons stained positive for γ-aminobutyric acid (GABA) and tyrosine hydroxylase (TH). In summary, we develop an experimental system for differentiating rES cells to tripotent neural progenitors, which may provide a powerful tool for pharmacological test and a valuable platform for studying the pathogenesis of many neurodegenerative disorders such as Parkinson's disease and the development of rat nervous system.
- Rat embryonic stem (rES) cells /
- Rat neural progenitor cells (rNPCs) /
- Neural differentiation /
- Neuron
These authors contributed equally to this work.
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