Generation of Regionally Specific Neural Progenitor Cells (NPCs) and Neurons from Human Pluripotent Stem Cells (hPSCs).
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.
School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ, 85287-9709, USA.
Josh
J
Cutts
School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ, 85287-9709, USA.
Nicholas
N
Brookhouser
School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ, 85287-9709, USA. David.Brafman@asu.edu.
David A
DA
Brafman
D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000235:genetics*; D006801:Humans; D058953:Neural Stem Cells / Q000166:cytology*; D009474:Neurons / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology
Anterior–posterior patterning; Disease modeling; Drug screening; Human pluripotent stem cells; Neural progenitor cells; Neurons; Regenerative medicine