title: Differentiation of serum-free embryoid bodies from human induced pluripotent stem cells into networks.
abstract: Three-dimensional aggregation cultures allow for complex development of differentiated human induced pluripotent stem cells. However, this approach is not easily amenable to live-cell imaging and electrophysiological applications due to the thickness and the geometry of the tissue. Here, we present an improvement on the traditional aggregation method by combining the use of cell culture inserts with serum-free embryoid bodies (SFEBs). The use of this technique allows the structures to maintain their three-dimensional structure while thinning substantially. We demonstrate that this technique can be used for electrophysiological recodings as well as live-cell calcium imaging combined with electrical stimulation, akin to organotypic slice preparations. This provides an important experimental tool that can be used to bridge 3-D structures with traditional monolayer approaches used in stem cell applications.
authors.affiliation: The New York Stem Cell Foundation Laboratory 178 Columbus Avenue #237064 New York, NY 10023, USA. mnestor@nyscf.org
authors.forename: Michael W
authors.initials: MW
authors.lastname: Nestor
authors.forename: Daniel
authors.initials: D
authors.lastname: Paull
authors.forename: Samson
authors.initials: S
authors.lastname: Jacob
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Sproul
authors.forename: Abdulaziz
authors.initials: A
authors.lastname: Alsaffar
authors.forename: Brian A
authors.initials: BA
authors.lastname: Campos
authors.forename: Scott A
authors.initials: SA
authors.lastname: Noggle
mesh_terms: D000328:Adult; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D004558:Electric Stimulation; D004566:Electrodes; D058732:Embryoid Bodies / Q000166:cytology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D008297:Male
chemical_list: D002118:Calcium