Differentiation of serum-free embryoid bodies from human induced pluripotent stem cells into networks

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.

► Developed a 3-D system for growing iPS cells based on organotypic slice culture ► Method induces the development of iPS cell-derived neurons and progenitors in vitro. ► SFEB Ca2 + signals can be imaged with electrical and chemical stimulation. ► Non-neuronal SFEBs from iPS-cells can be used for calcium imaging as well.