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In light of recent developments, artificially synthesized carbon\u2010based biomaterials such as carbon nanotubes and graphene have demonstrated feasibility in supporting stem cell attachment and differentiation. However, the applicability is significantly hampered by evidence of nanotoxic effects on multiple cell types. Thus, an emergent drive for an innovative carbonaceous biomaterial calls for a safer platform with comparable advantageous characteristics. Here, we showed for the first time, a natural coal\u2010based activated charcoal (AC) composite biosubstrate can support and promote neuronal differentiation in hESCs. The bio\u2010friendly AC composite biomatrices resulted in more matured neuron\u2010like cells. Both of axonal length and density were at least twice as long and abundant, respectively, when compared with control groups. A functional assay demonstrated that the derived neuron\u2010like cells responded to depolarization\u2010dependent synaptic recycling and may contain active synapses. In addition, the AC composite substrate can serve to concentrate growth factors and cell adhesion proteins, further encouraging attachment and hESC differentiation. Moreover, the AC composite biomaterial can potentially be economically manufactured as implantable three\u2010dimensional bioscaffolds, facilitating the regeneration of damaged neural and other tissues. \u00a9 2012 Wiley Periodicals, Inc. 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