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Tissue engineering, in the fields of skin regeneration,seeks to overcome the limitationsassociated with the use of immediate auto-grafts,since choosing the donor region of the patientconstitutes a risk for the patient himself and isinsufficient if the injury that has to be repairedis very large. The use of the pig’s small intestinesubmucosa (SIS) has being proved as a fillingbiomaterial to treat injuries, because of its specialcomposition, it lowers pain and inflammationsince its first application and it favours earlymobility to the wounded area. The present studydeveloped a protocol for the primary cultureof human keratinocytes from infant foreskins,and used small intestinal submucosa (SIS) likeculture substrate. Cellular adherence potentialand proliferation capability of the keratinocytesover this substrate was evaluated.