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Abstract Human pluripotent stem cell (hPSC)-derived cardiomyocytes provide a promising regenerative cell therapy for cardiovascular patients and an important model system to accelerate drug discovery. However, cost-effective and time-efficient platforms must be developed to evaluate the quality of hPSC-derived cardiomyocytes during biomanufacturing. Here, we develop a non-invasive label-free live cell imaging platform to predict the efficiency of hPSC differentiation into cardiomyocytes. Autofluorescence imaging of metabolic co-enzymes is performed under varying differentiation conditions (cell density, concentration of Wnt signaling activator) across five hPSC lines. Live cell autofluorescence imaging and multivariate classification models provide high accuracy to separate low (< 50%) and high (≥ 50%) differentiation efficiency groups (quantified by cTnT expression on day 12) within 1 day after initiating differentiation (area under the receiver operating characteristic curve, 0.91). This non-invasive and label-free method could be used to avoid batch-to-batch and line-to-line variability in cell manufacturing from hPSCs.
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Label-free optical imaging probing of metabolic states during human pluripotent stem cell differentiation to definitive endoderm
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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- PAR ID:
- 10488065
- Publisher / Repository:
- SPIE Photonics West
- Date Published:
- Format(s):
- Medium: X
- Location:
- San Francisco, CA
- Sponsoring Org:
- National Science Foundation
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