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Embryonic stem cells (ESC) are pluripotent, with the potential to differentiate into multiple cell types, making them a valuable tool for regenerative medicine and disease therapy. However, common culture methods face challenges, including strict operating procedures and high costs. Currently, Leukemia inhibitory factor (LIF), an indispensable bioactive protein for ESC culture, is typically applied to maintain self-renewal and pluripotency, but its instability and high cost limit its effectiveness in stable culture conditions. Hence, we have developed an innovative strategy using a soluble nanomaterial, metal-organic polyhedra (MOPs), to effectively maintain the self-renewal and pluripotency of ESC. The selected amino-modified vanadium-based MOP not only exhibits excellent biocompatibility and high stability but also possesses similar or even superior biological functions compared to commercial LIF. Due to the precise structure of MOPs, the active site responsible for maintaining ESC pluripotency has been identified and regulated at the molecular level. The new ESC culture method significantly reduces costs, simplifies preparation, and enhances the practicality of biopharmaceutical preparation and storage. This represents the first case of using MOPs to maintain self-renewal of ECS, opening an avenue for introducing advanced materials into the development of innovative ESC culture methods. Subject terms: Biomaterials - cells, Chemical biology

Endometrial receptivity is a critical determinant of embryo implantation and early pregnancy success; however, current methods for assessing endometrial receptivity remain poorly validated and insufficiently reliable for clinical application. Here, we establish a patient-derived vascularised endometrium-on-a-chip (EoC), successfully replicating the dynamic microenvironment and both temporal and spatial architecture of native endometrial tissue. Using our EoC, we develop a clinically relevant endometrial receptivity scoring system, ERS2, which integrates molecular profiling of established receptivity markers with quantitative analyses of angiogenesis. The ERS2 enables personalised assessment of endometrial health and implantation potential, addressing inter-patient variability often overlooked by conventional techniques. By leveraging our EoC to therapeutic monitoring, we observe progressive restoration of the endometrial microenvironment following platelet-rich-plasma treatments, highlighting the translational utility of our model. This study represents the innovative application of a patient-derived EoC and scoring system to assess receptivity, offering personalised infertility management and advancing targeted therapies in reproductive medicine. Accurate assessment of endometrial receptivity remains a challenge in infertility care. Here, authors present a patient-derived vascularised endometrium-on-a-chip and a scoring system for receptivity evaluation.

Oxygen tension is a key regulator of early human neurogenesis; however, quantifying intra-tissue O2 in 3D models for an extended period remains difficult. Existing approaches, such as needle-type fiber microsensors and intensity-based oxygen probes or time-domain lifetime imaging, either perturb the organoids or require high excitation doses that limit the measurement period. Here, we present a step-by-step protocol to measure intra-organoid oxygen in human cerebral organoids (hCOs) using embedded ruthenium-based CPOx microbeads and widefield frequency-domain fluorescence lifetime imaging microscopy (FD-FLIM). The workflow covers dorsal/ventral cerebral organoid patterning, organoid fusion at day 12 with co-embedded CPOx beads, standardized FD-FLIM acquisition (470-nm external modulation, 16 phases at 50 kHz, dual-tap camera), automated bead detection and lifetime extraction in MATLAB, and session-matched Stern–Volmer calibration with Ru(dpp)3(ClO4)2 to convert lifetimes to oxygen concentration. The protocol outputs per-bead oxygen maps and longitudinal patterns stratified by bead location (intra-organoid vs. gel) and sample state (healthy vs. abnormal), enabling direct linkage between developmental growth and oxygen dynamics. Key features • End-to-end workflow linking hCOs generation, on-gel bead embedding, and FD-FLIM oxygen readout.• Longitudinal single-organoid tracking of oxygen tension with bead-level metadata.• Reference-based lifetime calibration and reproducible camera/LED settings.• Ready-to-reuse materials, recipes, timing, and analysis logic.