The efficient transition of hPSCs to definitive endoderm (DE) progeny is an essential step toward disease modeling and the manufacturing of a wide range of cellular therapeutics in medical relevant quantities. Two-photon excited fluorescence (TPEF) imaging, as a non-invasive, non-destructive, label-free modality for metabolic studies, reveals the distinct metabolic switches during DE differentiation in a real-time monitoring mode. Since metabolic pathways orchestrate important regulatory mechanisms that influence and determine cell fate decisions, TPEF imaging serves as an important enabling technology in hPSC-based tissue engineering applications affording non-invasive determination of metabolic biomarkers and informing optimizations of hPSCs differentiation processes.
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