Establishment of a diverse head and neck squamous cancer cell bank using conditional reprogramming culture methods

Abstract

Most laboratory models of head and neck squamous cell cancer (HNSCC) rely on established immortalized cell lines, which carry inherent bias due to selection and clonality. We established a robust panel of HNSCC tumor cultures using a "conditional reprogramming" (CR) method, which utilizes a rho kinase inhibitor (Y-27632) and co-culture with irradiated fibroblast (J2 strain) feeder cells to support indefinite tumor cell survival. Sixteen CR cultures were successfully generated from 19 consecutively enrolled ethnically and racially diverse patients with HNSCC at a tertiary care center in the Bronx, NY. Of the 16 CR cultures, 9/16 were derived from the oral cavity, 4/16 were derived from the oropharynx, and 3/16 were from laryngeal carcinomas. Short tandem repeat (STR) profiling was used to validate culture against patient tumor tissue DNA. All CR cultures expressed ΔNp63 and cytokeratin 5/6, which are markers of squamous identity. Human papillomavirus (HPV) testing was assessed utilizing clinical p16 staining on primary tumors, reverse transcription polymerase chain reaction (RT-PCR) of HPV16/18-specific viral

Identification of HPV oncogene and host cell differentiation associated cellular heterogeneity in cervical cancer via single‐cell transcriptomic analysis

“…Lastly, precise processes of HPV life cycle and oncogenesis still remain largely unknown due to the lack of natural systems for HPV productive and abortive life cycles. It would be important to generate more patient‐derived clinically relevant cell models 37,41,42,72 …”

“…The precise process of HPV infection and oncogenesis can be experimentally determined and validated on patient-derived clinically relevant models, thus, making such a model is critically needed for studies of HPV biology and oncogenesis. Our group and others [36][37][38][39][40][41] recently established a clinically relevant cellular model with patient-derived, naturally infected HPVs, 3D ALI cultures from these PCDs employ the HPV life cycle and have been used for anti-HPV screening. 37,42 Long-term HPV/host interactions, especially proliferation manipulated by the interactions, become at high risk for the transformation of the host cells, which is not a primary intent of HPV infection.…”

CDK4/6 Inhibition Induces Senescence and Enhances Radiation Response by Disabling DNA Damage Repair in Oral Cavity Squamous Cell Carcinoma

“…Conditionally reprogrammed ‘CR’ cells were assigned cell line number during tumor harvesting. CR-06 and CR-018 are conditionally reprogrammed OCSCC CR cultures that have been previously described [ 38 ].…”

Characterization of a Diverse Set of Conditionally Reprogrammed Head and Neck Cancer Cell Cultures

“…Tumor tissue processing methods have been previously published, including specific media components. 24,28 Briefly, samples were rinsed with 100% ethanol and sterile phosphatebuffered saline (PBS) prior to placement in iced transport media. In some cases, tumor samples were stored in freezing media prior to processing, depending on lab workflow.…”

“…We have previously reported our initial experience using these methods. 28 Here, we describe the expansion of our CR culture library and further characterize the successfully cultured cells.…”