Irregular defects at sites of degenerative cartilage often accompany osteoarthritis (OA). The development of novel cell-/biomaterial-based cartilage tissue engineering methods to address these defects may provide a durable approach to hinder the development of OA. In this study, we fabricated a neocartilage patch by fusing cell aggregates onto a biodegradable nanofiber film for degenerative cartilage repair. Human mesenchymal stem/stromal cell (MSC) aggregates were prepared and induced for chondrogenesis in a thermosensitive hydrogel, poly (N-isopropylacrylamide-co-acrylic acid (p(NIPAAm-AA)). Cell migration mediated the formation of cell aggregates in the thermosensitive hydrogel and led to a cell-dense hollow shell structure. The chondrocytes derived from MSC aggregates in the hydrogel were evidenced by the expression of chondrogenesis-related genes and extracellular matrices. They were fused onto an electrospun film by mechanical force and spatial confinement to generate a neo-cartilage patch. The fabricated neocartilage patches may be able to integrate into the irregular defects under compressive stresses and achieve cartilage regeneration in vivo. Impact statement The formation of human mesenchymal stem/stromal cells aggregates in thermosensitive hydrogels was mechanistically examined. These in situ formed cell aggregates with enhanced chondrogenesis were bioengineered into a neocartilage patch for regeneration of superficial irregular cartilage defects.