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

ABSTRACT: 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.

ORGANISM(S): Homo sapiens

PROVIDER: GSE294833 | GEO | 2025/12/13

REPOSITORIES: GEO

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Project description:A reliable suspension-based platform for scaling engineered cardiac tissue (ECT) production from human induced pluripotent stem cells (hiPSCs) is crucial for regenerative therapies. Here, we compared the production and functionality of ECTs formed using our scaffold-based, engineered tissue microsphere differentiation approach with those formed using the prevalent scaffold-free aggregate platform. We utilized a microfluidic system for the rapid (1 million cells/min), high density (30, 40, 60 million cells/ml) encapsulation of hiPSCs within PEG-fibrinogen hydrogel microspheres. HiPSC laden-microspheres and aggregates underwent suspension-based cardiac differentiation in chemically defined media. In comparison to aggregates, microspheres maintained consistent size and shape initially, over time, and within and between batches. Initial size and shape coefficients of variation for microspheres were eight and three times lower, respectively, compared to aggregates. On day 10, microsphere cardiomyocyte (CM) content was 27% higher and the number of CMs per initial hiPSC was 250% higher than for aggregates. Contraction and relaxation velocities of microspheres were four and nine times higher than those of aggregates, respectively. Microsphere contractile functionality also improved with culture time, whereas aggregate functionality remained unchanged. Additionally, microspheres displayed improved β-adrenergic signaling responsiveness and uniform calcium transient propagation. Transcriptomic analysis revealed that while both microspheres and aggregates demonstrated similar gene regulation patterns associated with cardiomyocyte differentiation, heart development, cardiac muscle contraction, and sarcomere organization, the microspheres exhibited more pronounced transcriptional changes over time. Taken together, these results highlight the capability of the microsphere platform for scaling up biomanufacturing of ECTs in a suspension-based culture platform.

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

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