Primary cell culture of head and neck cancer: a challenge

Financial support: None. Conflicts of interest: No conflicts of interest declared concerning the publication of this article. Submitted: November 25, 2017. Accepted: July 13, 2018. The study was carried out at School of Pharmaceutical Sciences of Ribeirao Preto and Division of Head and Neck Surgery, Ribeirao Preto Medical School, University of Sao Paulo (USP), Ribeirao Preto, SP, Brazil abstract 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.