Primary cell culture of head and neck cancer: a challenge

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.

Ex Vivo Culture Models to Indicate Therapy Response in Head and Neck Squamous Cell Carcinoma

“…Furthermore, primary cells have a limited lifespan and are more sensitive to environmental changes and stress compared to immortalized cell lines. In addition, primary cultures are prone to microbial contaminations with bacteria and/or fungi, especially when the tissue is derived from locations with an extensive microbiome, such as the intestines or oral cavity [84,85]. Contamination with fibroblasts could also be a practical challenge, since fibroblasts are able to overgrow the culture because of their high proliferation rate [86,87].…”