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Holotomography has great potential in cell therapy, especially in the chimeric antigen receptor T cells (CAR-T) treatment. This technology provides valuable insights into cell behavior, quality, and function in both cell therapy research and clinical settings and goes a long way toward improving the clinical outcomes of cell-based therapy.

In cell therapy, HT enables the detailed visualization and characterization of cell morphology, viability, and distribution. This provides important information necessary for quality control and optimization of cell manufacturing processes. In addition, it can monitor cell behavior in real time, which helps to understand cell division, differentiation, and interaction with the surrounding environment, and to determine culture conditions for the optimal therapeutic effect. The quality and function of cells can be evaluated in a non-invasive and unlabeled manner, enabling fast and accurate quality control. This ensures consistency in cell therapy products.

In CAR-T treatment, CAR-T cell efficacy can be assessed, and treatment outcomes can be predicted by visualizing CAR-T cell morphology, active state, and interaction with target cells. In addition, by monitoring the expansion and proliferation of CAR-T cells in real time, the manufacturing process can be optimized to obtain the desired cell yield. The function of CAR-T cells can also be evaluated, which can be used to analyze target cell recognition, killing efficiency, cytokine secretion, etc., and improve CAR-T cell engineering strategies to improve treatment efficacy.

The utilization of HT provides valuable insights into cell behavior, quality, and function in cell therapy research and clinical settings, and greatly helps to improve the clinical outcomes of cell-based therapy.

Observing living stem cells is critical for understanding their behavior and unlocking their therapeutic potential. By studying stem cells in their native state, researchers can assess their pluripotency and potential for differentiation, laying the groundwork for various applications in regenerative medicine and tissue engineering.

Additionally, monitoring living stem cells over time provides valuable insights into their dynamic interactions with the surrounding microenvironment and their response to external stimuli. This knowledge is instrumental in advancing our understanding of stem cell biology and harnessing its therapeutic potential for treating various diseases and injuries.

In a 2021 paper published in Molecules and Cells, researchers demonstrated that normal cells and stem cells can be distinguished based on their refractive index (RI) and volume in HT images, eliminating the need for fluorescent markers (Kim et al., 2021).

This method effectively differentiated stem cells using RI and volume metrics, allowing quantification of nucleoli and vesicular structures unique to each cell type. Notably, stem cells exhibited significantly higher RI in vesicular structures compared to fibroblasts, as revealed by electron microscopy and HT analysis. These findings highlight HT’s potential to differentiate living stem cells from normal cells without fluorescence or specific biomarkers.

Red blood cells (RBCs) research in regenerative medicine focuses on harnessing their unique properties for therapeutic purposes, particularly in tissue repair and regeneration. While RBCs are primarily recognized for their role in oxygen transport, they also possess characteristics that make them promising candidates for regenerative medicine applications.

Quantitative analysis of RBCs using holotomography offers a comprehensive approach to understanding their properties in the context of regenerative medicine. In a study published in Stem Cell Reports, researchers utilized HT to evaluate differentiated RBC characteristics derived from human induced pluripotent stem cells (hiPSC) (Sivalingam et al., 2021).

The 3D refractive index (RI) distributions of purified enucleated RBCs were obtained through HT, enabling the quantification of various morphological, biochemical, and mechanical properties. Morphological characteristics such as cellular volume, surface area, and sphericity were determined from reconstructed 3D RI tomograms.

Additionally, cellular hemoglobin concentration (CHC) and content (CH) were calculated based on the mean RI of the cell and the known refraction increment of hemoglobin. This information offered insights into the hemoglobin content of individual RBCs, essential for understanding their oxygen-carrying capacity and therapeutic potential. Dynamic mechanical properties of RBCs, specifically membrane fluctuations, were also assessed using HT. By generating 2D membrane fluctuation maps and analyzing temporal standard deviations of mean cell height profiles, researchers quantify the spatial average of membrane fluctuation over the projected cell area. This parameter provides valuable insights into the mechanical flexibility and integrity of RBC membranes, crucial for their function in circulation and potential involvement in regenerative processes.

Overall, HT-based analysis of RBCs offers a comprehensive understanding of their structural, biochemical, and mechanical properties, providing valuable insights for regenerative medicine applications. This approach enables the characterization of RBCs at a single-cell level, facilitating the exploration of their potential therapeutic roles in tissue repair and regeneration.

White blood cells (WBCs) are integral to the immune system, defending against infections and diseases. The analysis of WBCs using HT technology holds promise for advancing regenerative medicine by enhancing our understanding of cell characteristics and interactions within the regeneration process. Such research endeavors are poised to contribute to the of innovative treatments leveraging the regenerative potential of the immune system.

A recent study utilized Holotomography to assess the functionality of WBCs separated through different methods, revealing the advantages of using microfluidic as the separation technique (Lee et al., 2023). HT allows visualization of the 3D structure and internal composition of cells, aiding in understanding traits like WBCs’ morphology, motility, and phagocytic ability.

Timelapse imaging at high temporal resolution enabled the monitoring of phagocyte dynamics, while HT-fluorescence correlative imaging allowed for the examination and comparison of the phagocytic ability based on different separation strategies. The study suggested that HT was an optimal tool that minimized cellular deformation for image analysis in regenerative research.