Poster #093, UC Davis Health System
Optimization of Expansion and Maturation of Human iPSC-Derived Astrocytes
Mentors: Celena Lozano, PhD Candidate and Kyle Fink, PhD
Human induced pluripotent stem cells (hiPSC)- derived models allow scientists to reprogram mature cells into pluripotent stem cells that can then be differentiated into multiple cell types including neurons, astrocytes, and cerebral organoids. These models provide a window into understanding human brain development and neurodevelopmental disorders. During this project, I gained experience culturing and replating NGN2-induced excitatory neurons, optimizing astrocyte culture conditions, and imaging and analyzing the size and shape of cerebral organoids. My primary project focused on comparing the survival, proliferation, morphology, and maturity of human astrocytes cultured in four conditions: (1) 10% FBS (2) serum-free media (3) serum free media with CNTF and (4) serum-free media with FGF2 and EGF. Astrocytes cultured in FBS had better survival and attachment, proliferated more rapidly, and exhibited a pancaked morphology. In comparison, the serum-free astrocytes had smaller and fewer surviving cells that proliferated slowly. Astrocytes cultured in serum-free media with different growth factors exhibited additional differences in morphology and proliferation. These findings demonstrate that culture conditions influence astrocyte morphology and growth. Optimizing astrocyte culture conditions is an important step towards developing physiologically-relevant human brain models which can improve our understanding of both typical neurodevelopment neurodevelopmental disorders.