TY - JOUR
T1 - Efficient and rapid derivation of primitive neural stem cells and generation of brain subtype neurons from human pluripotent stem cells
AU - Yan, Yiping
AU - Shin, Soojung
AU - Balendu, Shekhar Jha
AU - Liu, Qiuyue
AU - Sheng, Jianting
AU - Li, Fuhai
AU - Zhan, Ming
AU - Davis, Janine
AU - Bharti, Kapil
AU - Zeng, Xianmin
AU - Rao, Mahendra
AU - Malik, Nasir
AU - Vemuri, Mohan C.
PY - 2013
Y1 - 2013
N2 - Human pluripotent stem cells (hPSCs), including human embryonic stem cells and human induced pluripotent stem cells, are unique cell sources for disease modeling, drug discovery screens, and cell therapy applications. The first step in producing neural lineages from hPSCs is the generation of neural stem cells (NSCs). Current methods of NSC derivation involve the time-consuming, laborintensive steps of an embryoid body generation or coculture with stromal cell lines that result in low-efficiency derivation of NSCs. In this study, we report a highly efficient serum-free pluripotent stem cell neural induction medium that can induce hPSCs into primitive NSCs (pNSCs) in 7 days, obviating the need for time-consuming, laborious embryoid body generation or rosette picking. The pNSCs expressed the neural stem cell markers Pax6, Sox1, Sox2, and Nestin; were negative for Oct4; could be expanded for multiple passages; and could be differentiated into neurons, astrocytes, and oligodendrocytes, in addition to the brain region-specific neuronal subtypes GABAergic, dopaminergic, and motor neurons. Global gene expression of the transcripts of pNSCs was comparable to that of rosette-derived and human fetal-derived NSCs. This work demonstrates an efficient method to generate expandable pNSCs, which can be further differentiated into central nervous system neurons and glia with temporal, spatial, and positional cues of brain regional heterogeneity. This method of pNSC derivation sets the stage for the scalable production of clinically relevant neural cells for cell therapy applications in good manufacturing practice conditions.
AB - Human pluripotent stem cells (hPSCs), including human embryonic stem cells and human induced pluripotent stem cells, are unique cell sources for disease modeling, drug discovery screens, and cell therapy applications. The first step in producing neural lineages from hPSCs is the generation of neural stem cells (NSCs). Current methods of NSC derivation involve the time-consuming, laborintensive steps of an embryoid body generation or coculture with stromal cell lines that result in low-efficiency derivation of NSCs. In this study, we report a highly efficient serum-free pluripotent stem cell neural induction medium that can induce hPSCs into primitive NSCs (pNSCs) in 7 days, obviating the need for time-consuming, laborious embryoid body generation or rosette picking. The pNSCs expressed the neural stem cell markers Pax6, Sox1, Sox2, and Nestin; were negative for Oct4; could be expanded for multiple passages; and could be differentiated into neurons, astrocytes, and oligodendrocytes, in addition to the brain region-specific neuronal subtypes GABAergic, dopaminergic, and motor neurons. Global gene expression of the transcripts of pNSCs was comparable to that of rosette-derived and human fetal-derived NSCs. This work demonstrates an efficient method to generate expandable pNSCs, which can be further differentiated into central nervous system neurons and glia with temporal, spatial, and positional cues of brain regional heterogeneity. This method of pNSC derivation sets the stage for the scalable production of clinically relevant neural cells for cell therapy applications in good manufacturing practice conditions.
KW - Astrocytes
KW - Cell culture
KW - Nestin
KW - Neural differentiation
KW - Neural induction
KW - Neural stem cell
KW - Neuron
KW - Oligodendrocytes
UR - http://www.scopus.com/inward/record.url?scp=84886528116&partnerID=8YFLogxK
U2 - 10.5966/sctm.2013-0080
DO - 10.5966/sctm.2013-0080
M3 - Article
C2 - 24113065
AN - SCOPUS:84886528116
SN - 2157-6564
VL - 2
SP - 862
EP - 870
JO - Stem Cells Translational Medicine
JF - Stem Cells Translational Medicine
IS - 11
ER -