Macroporous networks of poly(ethylene glycol) (PEG) with interconnected pores can be created by cryogelation techniques. In this study, we describe the potential application of such PEG cryogels as scaffolds for cartilage tissue engineering. Three-dimensional macroporous cryogels were evaluated for chondrocyte growth and production of cartilage-specific extracellular matrix (ECM). Seeded primary bovine chondrocytes showed homogeneous distribution throughout the cryogels. DNA content suggests continuous cell proliferation over 4 weeks of in vitro culture. Analysis of the composition of cell-secreted ECM showed a culture-time-dependent increase in the amount of glycosaminoglycan and collagen. The production of ECM by chondrocytes was confirmed using scanning electron microscopy analysis. Further histological and immunohistological analysis of the cell-laden scaffold confirmed the presence of accumulated cartilage-specific ECM within the scaffold. The interconnected macroporous network promoted diffusion of cell-secreted matrix within the cryogels. Our results indicated that interconnected macroporous PEG cryogels successfully supported attachment, viability, proliferation, and biosynthetic activity of seeded chondrocytes.