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As mechanical properties of cell culture substrates matter, methods for mechanical characterization of scaffolds on a relevant length scale are required. We used multiple particle tracking microrheology to close the gap between elasticity determined from bulk measurements and elastic properties sensed by cells. Structure and elasticity of macroporous, three-dimensional cryogel scaffolds from mixtures of hyaluronic acid (HA) and collagen (Coll) were characterized. Both one-component gels formed homogeneous networks, whereas hybrid gels were heterogeneous in terms of elasticity. Most strikingly, local elastic moduli were significantly lower than bulk moduli presumably due to non-equilibrium chain conformations between crosslinks. This was more pronounced in Coll and hybrid gels than in pure HA gels. Local elastic moduli were similar for all gels, irrespective of their different swelling ratio and bulk moduli. Fibroblast cell culture proved the biocompatibility of all investigated compositions. Coll containing gels enabled cell migration, adhesion and proliferation inside the gels.