TY - CHAP
T1 - Generation of regionally specific neural progenitor cells (NPCs) and neurons from human pluripotent stem cells (HPSCs)
AU - Cutts, Josh
AU - Brookhouser, Nicholas
AU - Brafman, David
PY - 2016
Y1 - 2016
N2 - Neural progenitor cells (NPCs) derived from human pluripotent stem cells (hPSCs) are a multipotent cell population capable of long-term expansion and differentiation into a variety of neuronal subtypes. As such, NPCs have tremendous potential for disease modeling, drug screening, and regenerative medicine. Current methods for the generation of NPCs results in cell populations homogenous for pan-neural markers such as SOX1 and SOX2 but heterogeneous with respect to regional identity. In order to use NPCs and their neuronal derivatives to investigate mechanisms of neurological disorders and develop more physiologically relevant disease models, methods for generation of regionally specific NPCs and neurons are needed. Here, we describe a protocol in which exogenous manipulation of WNT signaling, through either activation or inhibition, during neural differentiation of hPSCs, promotes the formation of regionally homogenous NPCs and neuronal cultures. In addition, we provide methods to monitor and characterize the efficiency of hPSC differentiation to these regionally specific cell identities.
AB - Neural progenitor cells (NPCs) derived from human pluripotent stem cells (hPSCs) are a multipotent cell population capable of long-term expansion and differentiation into a variety of neuronal subtypes. As such, NPCs have tremendous potential for disease modeling, drug screening, and regenerative medicine. Current methods for the generation of NPCs results in cell populations homogenous for pan-neural markers such as SOX1 and SOX2 but heterogeneous with respect to regional identity. In order to use NPCs and their neuronal derivatives to investigate mechanisms of neurological disorders and develop more physiologically relevant disease models, methods for generation of regionally specific NPCs and neurons are needed. Here, we describe a protocol in which exogenous manipulation of WNT signaling, through either activation or inhibition, during neural differentiation of hPSCs, promotes the formation of regionally homogenous NPCs and neuronal cultures. In addition, we provide methods to monitor and characterize the efficiency of hPSC differentiation to these regionally specific cell identities.
KW - Anterior–posterior patterning
KW - Disease modeling
KW - Drug screening
KW - Human pluripotent stem cells
KW - Neural progenitor cells
KW - Neurons
KW - Regenerative medicine
UR - http://www.scopus.com/inward/record.url?scp=84984998266&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84984998266&partnerID=8YFLogxK
U2 - 10.1007/7651_2016_357
DO - 10.1007/7651_2016_357
M3 - Chapter
C2 - 27106497
AN - SCOPUS:84984998266
VL - 1516
T3 - Methods in Molecular Biology
SP - 121
EP - 144
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -