Background: Skin is a 3-dimensional (3-D) tissue that mainly consists 2 layers, comprising the epidermis and dermis. Skin tissue engineering scaffolds are used commonly as 3-D analogs of the extracellular matrix (ECM) of the skin. Bacterial cellulose (BC) has great importance in skin tissue engineering because of its resemblance to ECM and its biocompatibility. The lack of 3-D microporosity and limited biodegradation capacity has restricted its application as a scaffold for skin tissue engineering. Controlled 3-D microporosity of BC via surface modification techniques are required for potential tissue engineering applications.Methods: Freeze-drying is an ex-situ surface modification technique for making macroporous BC scaffolds (MBCSs). This study proposed a new approach to the freeze-drying method for the arrangement of the pore size of MBCSs specifically for the human keratinocyte cell line (KER-CT). Different concentrations of MBCS (0.25%, 0.50%, and 0.75%) were prepared and the KER-CT cell viability was detected via 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay.Result: The results of this study indicated that the KER-CT cells were able to proliferate all of the concentrations of MBCS, and the best cell viability value was observed with 0.75% MBCS. The results were supported by FESEM and light microscopic observations.Conclusion: These findings suggested that 0.75% MBCS might be of use in epidermal tissue engineering applications