Electro-assisted cell printing directs in vitro gastrulation and epiblast-like morphogenesis from human pluripotent stem cells II

This study develop a versatile techonology to largely generate biomimetic epiblast-like tissues based on human induced pluripotent stem cells (hiPSCs) and well-defined biomatrix. Single hiPSCs are wrapped in laminin-alginate (L-A) hydrogel with optimized concentration and stably fabricated a large number of uniform hiPSCs-contained microspheres via electro-assisted bioprinting technology. It was observed that hiPSCs self-organized into disc-like cell clusters in L-A microspheres. Transition of microenvironment to L-A microspheres triggers the fate specification of disc-shaped tissues towards gastrulation with the identity resemble posterior epiblast population of human embryos around gastrulating stage, which is supported by transcriptome analysis. Overall design: To investigate the time-dependent change of the cell fate of hiPSC clusters within bioprinted microspherese. We set samples rep1/rep2 of cell clusters on day6 in the persistent presence of blebbistatin. And samples rep1/2 of denoting the sample named Day6_dif_1 and presenting the differentiation of cell clusters on day6 for 4 days.