Composite multicellular spheroids composed of mesenchymal stem cells (MSCs) and synthetic biodegradable nanofilaments are fabricated. Extracellular‐matrix‐mimicking nanofilaments, prepared from transverse fragmentation of semicrystalline poly(L‐lactic acid) nanofibers and subsequent surface modification with cell adhesive peptides, are used to form composite multicellular spheroids with MSCs by cellular self‐assembly. The size of the composite spheroids could be readily controlled with the integrated amount of the nanofilaments. The composite spheroids show enhanced adipogenic potential compared to homotypic spheroids. The resultant spheroids are used as building blocks for 3D biohybrid construction with the assistance of a microstructured scaffold fabricated by a direct polymer melt deposition process. An angiogenic growth factor, basic fibroblast growth factor, is also locally delivered in a sustained fashion from the heparinized scaffold surface for facile neovascularization of adipogenic tissue. The produced multiscaled and multifunctional hybrid MSC construct enable the successful formation of vascularized adipose tissue in vivo.
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