Introduction: Head and neck cancer presents a high rate of recurrence and mortality, considering the sites affected. The use of primary culture allows pre-clinical trials that would not be possible in humans or would require a long time until the initial tests were approved. Objective: To establish primary culture of carcinomas and the disease‐free surgical margin of individuals affected by neck cancer. Methods: Fragments of 6 cases of oral cavity carcinoma and 2 cases of non-malignant tissue (surgical margin) of patients with oral cancer were collected immediately after surgical resection. These specimens were packed in complete DMEM (Dulbecco Modified Eagle’s Medium, SIGMA  ) supplemented with 10% inactivated Bovine Fetal Serum (BFSi) and 5% antibiotic / antimycotic - and kept on ice for transportation to the Molecular Marker Laboratory And Cancer Cell Signaling in FCFRP-USP. Processing was carried out in a biosafety booth in a cell culture room, 2 hours after collection maximum. All specimens collected were advanced tumors of the oral cavity. Results: From the 6 cases collected and kept in culture, only 2 presented uncontrollable bacterial contamination and were discarded. Two other cases released fibroblasts in the first 3 to 5 days and the observation of neoplastic cells (keratinocytes) was only possible after seven to ten days. Both cell types exhibited monolayer expansion. Conclusion: The use of explants to establish the initial stages of primary culture of head and neck cancer is a viable and easily reproducible alternative. The effective success rate is achieved in 20-30% of the cases and the control of the contamination presents itself as one of the biggest obstacles to be surpassed in the initial stages of cultivation.