Natural extracellular matrices often have a filamentous nature, however, only a limited number of artificial extracellular matrices have been designed from nanofibrillar building blocks. Here we report the preparation of temperature-responsive nanofibrillar hydrogels from rod-shaped cellulose nanocrystals (CNCs) functionalized with a copolymer of N-isopropylacrylamide and N,N'-dimethylaminoethyl methacrylate. The composition of the copolymer was tuned to achieve gelation of the suspension of copolymer-functionalized CNCs at 37 °C in cell culture medium and gel dissociation upon cooling it to room temperature. The mechanical properties and the structure of the hydrogel were controlled by changing copolymer composition and the CNC-to-copolymer mass ratio. The thermoreversible gels were used for the encapsulation and culture of fibroblasts and T cells and showed low cytotoxicity. Following cell culture, the cells were released from the gel by reducing the temperature, thus, enabling further cell characterization. These results pave the way for the generation of injectable temperature-responsive nanofibrillar hydrogels. The release of cells following their culture in the hydrogels would enable enhanced cell characterization and potential transfer in a different cell culture medium.