Work

Label-free method to monitor metabolism during long-term culture of human pluripotent stem cell derived cardiomyocytes

Year: 2025

Type: article

Abstract: Significance: Human pluripotent stem-cell-derived cardiomyocytes (hPSC-CMs) are a powerful tool for drug discovery, and metabolic changes are associated with their long-term culture and maturation. However, the lack of technologies to monitor hPSC-derived cardiomyocyte metabolism during long-term culture presents a major technical bottleneck. Aim: metabolic maturation of hPSC-CMs are limited by traditional assessment methods, which are generally time-consuming, destructive to samples, and lack single-cell resolution. We report a rapid, noninvasive imaging-based method to monitor hPSC-CM metabolism throughout extended culture (90+ days). Approach: Label-free optical metabolic imaging (OMI) of autofluorescent metabolic coenzymes was performed at multiple time points during the extended culture maturation process. In addition, OMI monitored hPSC-CMs grown on substrates with varying stiffness and on cardiomyocytes derived from induced pluripotent stem cells associated with cardiac arrhythmia. OMI was paired with immunofluorescence to validate structural maturation. Results: culturing. It can also detect metabolic differences induced by substrates of varying stiffnesses, can distinguish diseased from normal cell lines, and is sensitive to patient-level metabolic heterogeneity. Conclusions: Our results demonstrate that label-free OMI can be used to monitor metabolic changes in hPSC-CMs under varying culture conditions in a rapid, non-destructive manner with single-cell resolution, providing insight into metabolic transitions arising from time in culture, culture conditions, or disease states.

Source: Biophotonics discovery.

DOI: 10.1117/1.bios.2.2.025001