RT Journal Article
SR Electronic
T1 Tracking connectivity maps in human stem cell–derived neuronal networks by holographic optogenetics
JF Life Science Alliance
JO Life Sci. Alliance
FD Life Science Alliance LLC
SP e202101268
DO 10.26508/lsa.202101268
VO 5
IS 7
A1 Felix Schmieder
A1 Rouhollah Habibey
A1 Johannes Striebel
A1 Lars Büttner
A1 Jürgen Czarske
A1 Volker Busskamp
YR 2022
UL https://www.life-science-alliance.org/content/5/7/e202101268.abstract
AB Neuronal networks derived from human induced pluripotent stem cells have been exploited widely for modeling neuronal circuits, neurological diseases, and drug screening. As these networks require extended culturing periods to functionally mature in vitro, most studies are based on immature networks. To obtain insights on long-term functional features, we improved a glia–neuron co-culture protocol within multi-electrode arrays, facilitating continuous assessment of electrical features in weekly intervals. By full-field optogenetic stimulation, we detected an earlier onset of neuronal firing and burst activity compared with spontaneous activity. Full-field stimulation enhanced the number of active neurons and their firing rates. Compared with full-field stimulation, which evoked synchronized activity across all neurons, holographic stimulation of individual neurons resulted in local activity. Single-cell holographic stimulation facilitated to trace propagating evoked activities of 400 individually stimulated neurons per multi-electrode array. Thereby, we revealed precise functional neuronal connectivity motifs. Holographic stimulation data over time showed increasing connection numbers and strength with culture age. This holographic stimulation setup has the potential to establish a profound functional testbed for in-depth analysis of human-induced pluripotent stem cell-derived neuronal networks.