TY - JOUR
T1 - Efficient derivation of sympathetic neurons from human pluripotent stem cells with a defined condition
AU - Kirino, Kosuke
AU - Nakahata, Tatsutoshi
AU - Taguchi, Tomoaki
AU - Saito, Megumu K.
N1 - Funding Information:
We thank Dr. A. Niwa for scientific comments, and Ms. Harumi Watanabe for providing administrative assistance. We also thank Dr. Peter Karagiannis for reading the manuscript. This work was supported by a grant for the Core Center for iPS Cell Research of Research Center Network for Realization of Regenerative Medicine from the Japan Agency for Medical Research and Development (AMED) [T.N. and M.K.S.]; the Acceleration Program for Intractable Diseases Research utilizing Disease-specific iPS cells from AMED [17935244 to M.K.S.]; the Japan Society for the Promotion of Science (JSPS) KAKENHI grant numbers 16673093 [M.K.S.] and 16H02682 [T.N. and T.T.], Mochida Memorial Foundation for Medical and Pharmaceutical Research [M.K.S.], Takeda Science Foundation [M.K.S.] and iPS Cell Research Fund [M.K.S.]. The authors declare no conflicts of interest.
Publisher Copyright:
© 2018, The Author(s).
PY - 2018/12/1
Y1 - 2018/12/1
N2 - Sympathetic neurons (SNs) are an essential component of the autonomic nervous system. They control vital bodily functions and are responsible for various autonomic disorders. However, obtaining SNs from living humans for in vitro study has not been accomplished. Although human pluripotent stem cell (hPSC)-derived SNs could be useful for elucidating the pathophysiology of human autonomic neurons, the differentiation efficiency remains low and reporter-based cell sorting is usually required for the subsequent pathophysiological analysis. To improve the efficiency, we refined each differentiation stage using PHOX2B::eGFP reporter hPSC lines to establish a robust and efficient protocol to derive functional SNs via neuromesodermal progenitor-like cells and trunk neural crest cells. Sympathetic neuronal progenitors could be expanded and stocked during differentiation. Our protocol can selectively enrich sympathetic lineage-committed cells at high-purity (≈80%) from reporter-free hPSC lines. Our system provides a platform for diverse applications, such as developmental studies and the modeling of SN-associated diseases.
AB - Sympathetic neurons (SNs) are an essential component of the autonomic nervous system. They control vital bodily functions and are responsible for various autonomic disorders. However, obtaining SNs from living humans for in vitro study has not been accomplished. Although human pluripotent stem cell (hPSC)-derived SNs could be useful for elucidating the pathophysiology of human autonomic neurons, the differentiation efficiency remains low and reporter-based cell sorting is usually required for the subsequent pathophysiological analysis. To improve the efficiency, we refined each differentiation stage using PHOX2B::eGFP reporter hPSC lines to establish a robust and efficient protocol to derive functional SNs via neuromesodermal progenitor-like cells and trunk neural crest cells. Sympathetic neuronal progenitors could be expanded and stocked during differentiation. Our protocol can selectively enrich sympathetic lineage-committed cells at high-purity (≈80%) from reporter-free hPSC lines. Our system provides a platform for diverse applications, such as developmental studies and the modeling of SN-associated diseases.
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U2 - 10.1038/s41598-018-31256-1
DO - 10.1038/s41598-018-31256-1
M3 - Article
C2 - 30150715
AN - SCOPUS:85052332509
SN - 2045-2322
VL - 8
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 12865
ER -
