Quantitative phase imaging with temporal kinetics predicts hematopoietic stem cell diversity
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Abstract
Innovative identification technologies for hematopoietic stem cells (HSCs) have advanced the frontiers of stem cell biology. However, most analytical techniques capture only a single snapshot, disregarding the temporal context. A comprehensive understanding of the temporal heterogeneity of HSCs necessitates live-cell, real-time and non-invasive analysis. Here, we developed a prediction system for HSC diversity by integrating single-HSC ex vivo expansion technology with quantitative phase imaging (QPI)-driven machine learning. By analyzing single-cell kinetics with QPI, we discovered previously undetectable diversity among HSCs that snapshot analysis fails to capture. Our QPI-driven algorithm quantitatively evaluates the stemness of individual HSCs and incorporates temporal information to significantly improve prediction accuracy. This platform marks a paradigm shift from “identification” to “prediction”, enabling us to forecast HSC status by analyzing their past temporal kinetics.
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between changes in Hlf-tdTomato expression and temporal differentiation in HSCs was confirmed (Fig. 2E,157Fig. S3, F and G). To determine whether Hlf-tdTomato expression levels act as a functional indicator of158HSCs, tdTomato high and low expanded HSCs were transplanted into irradiated recipients, and long-term159bone marrow reconstitution abilities were evaluated. HSCs with high tdTomato demonstrate robust long-160term marrow reconstitution capabilities. Conversely, chimerism decreased in those who received161tdTomato-low HSCs (Fig. 2, F to H). Furthermore, the analysis of the spatiotemporal dynamics of early162hematopoiesis post-transplantation revealed that tdTomato high HSCs gradually reconstituted systemic163hematopoiesis, whereas tdTomato low HSCs induced rapid hematopoiesis between day 14 and 21 (Fig.1642, I to L).
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between changes in Hlf-tdTomato expression and temporal differentiation in HSCs was confirmed (Fig. 2E,157Fig. S3, F and G). To determine whether Hlf-tdTomato expression levels act as a functional indicator of158HSCs, tdTomato high and low expanded HSCs were transplanted into irradiated recipients, and long-term159bone marrow reconstitution abilities were evaluated. HSCs with high tdTomato demonstrate robust long-160term marrow reconstitution capabilities. Conversely, chimerism decreased in those who received161tdTomato-low HSCs (Fig. 2, F to H). Furthermore, the analysis of the spatiotemporal dynamics of early162hematopoiesis post-transplantation revealed that tdTomato high HSCs gradually reconstituted systemic163hematopoiesis, whereas tdTomato low HSCs induced rapid hematopoiesis between day 14 and 21 (Fig.1642, I to L).
This is a really nice demonstration that the levels of Hlf-tdTomato are indicators of 'stemness' but it would also be nice to see a confirmation that the fusion protein is expressed at levels (or not) that are similar to Hlf. Either would be fine given it is a clear indicator of 'stemness' but the molecular correlation would provide clearer ties to the mechanism of 'stemness.'
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