Presentation Information
[3FMBS-08]Photoactivatable PEG lipid platform for spatiotemporally controlled single-cell manipulation
○Shinya Yamahira1, Satoshi Yamaguchi1,2 (1. The University of Osaka (Japan), 2. The University of Tokyo (Japan))
Keywords:
Photoactivatable PEG lipid,Single-cell manipulation,Light-controlled cell capture,Immune–cancer cell interaction,Circulating tumor cells (CTCs)
[Purpose]
Recent advances in single-cell technologies have demonstrated that cellular heterogeneity plays essential roles in immune activation, cancer metastasis, and tissue development. Conventional bulk assays average signals from large cell populations and are therefore insufficient to resolve diverse cellular responses. Technologies enabling precise manipulation, isolation, and analysis of individual cells are thus increasingly important for biological and biomedical research. Surface-based approaches that require only simple coating processes, rather than complex microfabricated structures, are particularly attractive because they can be readily applied to diverse materials and experimental platforms. To address these challenges, we developed a photoactivatable polyethylene glycol (PEG) lipid platform for spatiotemporally controllable single-cell manipulation.
[Method]
PEG lipid consists of a hydrophilic PEG chain and a hydrophobic lipid anchor; the anchor interacts with cell membranes. Thus, coating solid substrates with PEG lipids enables immobilization of a wide variety of cells on the surface. To achieve light-controlled cell capture, we attached a second lipid chain via a photocleavable linker to the PEG lipid. This design renders the molecule inactive prior to irradiation, as the two hydrophobic lipid chains intermolecularly aggregate on the surface, hindering access of the lipid anchors to the cell membrane. Light irradiation cleaves one of the lipid chains, reducing hydrophobicity and resolving the aggregation, which exposes the remaining lipid anchor and enables cell capture. As a result, cells seeded on this surface are captured only in the irradiated regions, allowing spatially controlled single-cell positioning.
[Results]
The light-activatable PEG-lipid interface was applied to construct controlled immune–cancer cell interaction systems. Sequential cycles of patterned irradiation, cell seeding, and washing enabled precise side-by-side arrangement of immune and cancer cells with single-cell resolution. This configuration supported high-throughput single-cell analysis of immune–cancer interactions through real-time observation of cytotoxic responses and morphological dynamics. Furthermore, the system was applied to isolate circulating tumor cells from mouse blood samples containing human colon cancer cells in a microfluidic channel. Target tumor cells were identified microscopically and immobilized by localized laser irradiation, while non-target blood cells were removed by washing.
[Conclusion]
Overall, the photo-activatable PEG lipid provides a simple and versatile platform for optically addressable manipulation of individual cells on various substrates. This technology is expected to serve as a platform for advanced single-cell studies in cell biology, diagnostics, and cancer immunotherapy research.
Recent advances in single-cell technologies have demonstrated that cellular heterogeneity plays essential roles in immune activation, cancer metastasis, and tissue development. Conventional bulk assays average signals from large cell populations and are therefore insufficient to resolve diverse cellular responses. Technologies enabling precise manipulation, isolation, and analysis of individual cells are thus increasingly important for biological and biomedical research. Surface-based approaches that require only simple coating processes, rather than complex microfabricated structures, are particularly attractive because they can be readily applied to diverse materials and experimental platforms. To address these challenges, we developed a photoactivatable polyethylene glycol (PEG) lipid platform for spatiotemporally controllable single-cell manipulation.
[Method]
PEG lipid consists of a hydrophilic PEG chain and a hydrophobic lipid anchor; the anchor interacts with cell membranes. Thus, coating solid substrates with PEG lipids enables immobilization of a wide variety of cells on the surface. To achieve light-controlled cell capture, we attached a second lipid chain via a photocleavable linker to the PEG lipid. This design renders the molecule inactive prior to irradiation, as the two hydrophobic lipid chains intermolecularly aggregate on the surface, hindering access of the lipid anchors to the cell membrane. Light irradiation cleaves one of the lipid chains, reducing hydrophobicity and resolving the aggregation, which exposes the remaining lipid anchor and enables cell capture. As a result, cells seeded on this surface are captured only in the irradiated regions, allowing spatially controlled single-cell positioning.
[Results]
The light-activatable PEG-lipid interface was applied to construct controlled immune–cancer cell interaction systems. Sequential cycles of patterned irradiation, cell seeding, and washing enabled precise side-by-side arrangement of immune and cancer cells with single-cell resolution. This configuration supported high-throughput single-cell analysis of immune–cancer interactions through real-time observation of cytotoxic responses and morphological dynamics. Furthermore, the system was applied to isolate circulating tumor cells from mouse blood samples containing human colon cancer cells in a microfluidic channel. Target tumor cells were identified microscopically and immobilized by localized laser irradiation, while non-target blood cells were removed by washing.
[Conclusion]
Overall, the photo-activatable PEG lipid provides a simple and versatile platform for optically addressable manipulation of individual cells on various substrates. This technology is expected to serve as a platform for advanced single-cell studies in cell biology, diagnostics, and cancer immunotherapy research.
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