Establishment and characterization of novel human oral squamous cell carcinoma cell lines from advanced‑stage tumors of buccal mucosa

Abstract

Oral squamous cell carcinoma (OSCC) is a leading cause of mortality in India owing to the high percentage of tobacco chewers, smokers and alcohol consumption. OSCC is highly heterogeneous in nature; therefore poses a challenge in the treatment of the patient. To better understand the heterogeneity of the tumors, an in vitro cell line model is required. However, the efficiency of establishing cell lines from the oral tumors is low. In the present study, three novel cell lines, namely ACOSC3, ACOSC4, and ACOSC16, were isolated and characterized from advanced-stage treatment-naive OSCCs originating from the buccal mucosa. The three cell lines exhibited polygonal morphology, which is typical of epithelial cells. Furthermore, immunofluorescence revealed the expression of keratins 8 and 14, thereby confirming the epithelial origin of the cells. DNA content analysis of the three OSCC cell lines revealed aneuploidy. Furthermore, an in vitro orosphere assay revealed the formation of primary orospheres. Notably, the OSCC cell lines were able to give rise to tumors when administered subcutaneously into non-obese diabetic/severe combined immune deficiency mice. The novelty of the cell lines was also validated by performing short tandem repeat profiling; the STR profiles of the present cell lines did not significantly match with any known established OSCC cell lines present in the DSMZ database, thereby confirming the unique identity of these lines. These cell lines established from tumor samples derived from Indian OSCC patients provide a valuable resource to understand the molecular mechanism involved in tumor resistance and recurrence.

Decellularized Extracellular Matrix Composite Hydrogel Bioinks for the Development of 3D Bioprinted Head and Neck in Vitro Tumor Models

“…In addition, our spheroids showed keratin expression after 19 days of culture comparable to the xenograft mouse model suggesting that the cellular function of the moderately differentiated squamous cell carcinoma cell lines was maintained. Keratin expression has been used as a method for HNSCC model validation both in 2D and 3D culture models. − …”

Establishment and characterization of novel autologous pair primary cultures from two Indian non-habitual tongue carcinoma patients

“…Various articles report the establishment of epithelial cell lines from patient samples. These epithelial cell lines were derived from different tissue origins, for example, buccal mucosa (15), gingivobuccal mucosa (14), oral cavity (30–32), tongue (13,33), sinonasal (34), pyriform fossa (35), lower alveolus and retromolar trigone (33). To elucidate the impact of different mutations on tumor behaviour, recently 16 cell lines from HNSCC were established by Hayes et.al (36).…”

“…Certain studies have also reported directly digesting the tumor tissue and seeding the cell suspension (30,33). Majority of reports have described establishing primary cultures by explant culture method (13–15,32,34). The current study also used explant culture method for MhCT12 and MhCT08 establishment.…”

“…There have been various reports of establishment of oral cancer cell lines from tumors (13–15). In USA and UK initial attempts were made to establish cell lines from head and neck squamous cell carcinoma (HNSCC) patients who had undergone chemo and radiotherapy (16,17).…”

Context-dependent effect of sPLA2-IIA induced proliferation on murine hair follicle stem cells and human epithelial cancer

“…We have used ACOSC3, ACOSC4 and ACOSC16 OSCC cell lines, which were recently established in our laboratory from the advanced stage treatment naïve buccal mucosa tumours [39]. For skin cancer, we have used the human squamous cell carcinoma (SCC) line A3886 obtained from Dr. Colin Jamora's Lab at Instem, Bengaluru [40].…”