Directing Human Induced Pluripotent Stem Cell-Derived Embryonic-like Organoid with 3D Biomimetic Cryogel Mechanical Microenvironment for Neural Induction.

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We aim to investigate how using cryogel sandwich model by embedding cell clusters with layer-by-layer cryogel could enhance mechanical constraints. First, we studied cryogel`s topology by using Hirox, and SEM microscope analysis revealed that cryogel surface topology consists of voids that appear as hill-valley-like structures composed of pockets like void space which can trap microbeads. Thus, cryogel facilitates spontaneous cell cohesion, leading to spherical-shaped aggregates within 24h. After cryogel embedding the aggregates acquire a teardrop-like shape. Eventually, on day 3 these aggregates showed up ovoid shape which resembles a symmetrical breaking process. Day 4 showed polarized aggregates with the posterior axis.