A hydrogel scaffold with a non-fouling but specific cancer cell-adhesive surface was fabricated through surface modification using β-cyclodextrin-based host-guest chemistry. Interestingly, the hydrogel surface not only selectively captured specific cancer cells, but also grew the cells into multicellular spheroids. The spheroids could be released without damaging the cell viability through replacing the host moieties on the scaffold, and the released spheroids showed no changes in size or morphology.