Formation of assembloids by DNA-mediated synthetic cell self-assembly

This study presents a bottom-up synthetic tissue engineering strategy in which individual droplet-supported lipid bilayer synthetic cells self-assemble into millimetre-scale 3D constructs through programmable DNA-mediated adhesion. Cholesterol-tagged single-stranded DNA anchors were inserted into the synthetic cell membrane and paired with complementary linker strands to drive inter-synthetic-cell binding, enabling the formation of free-floating constructs, multi-zone assemblies, and standardized well-plate-format synthetic tissues. By varying DNA strand length and surface density, the authors encoded spatial organization and tuned emergent mechanical properties of the resulting constructs, while also showing that the system remains structurally stable in serum-supplemented cell culture medium and can be reversibly disassembled by DNA strand displacement. The platform was further functionalized with T cell-stimulatory antibodies to generate synthetic tissues with distinct reaction zones resembling aspects of lymph node organization. Primary human CD8+ T cells infiltrated these assembloids, formed proliferation clusters within the 3D structures, and showed activation profiles comparable to Dynabeads controls, including approximately 40% CD25-positive cells. Overall, the work establishes a controllable, modular and reversible synthetic-cell-based assembloid platform for tissue-mimetic culture systems and immune cell engineering.