sepharose and Mouth-Neoplasms

A perfusion three-dimensional (3D) cell culture microfluidic chip has been developed for real-time and non-invasive impedimetric monitoring of cell proliferation and chemosensitivity. In this study, human oral cancer cells (OEC-M1) were encapsulated in 3D agarose scaffold and cultured in a miniaturized chamber under perfusion of tested substance. This setting provides a more in vitro physiologically relevant microenvironment to better mimic the complex in vivo microenvironment. A pair of vertical electrodes was embedded at the opposite sidewalls of the culture chamber for the on-site impedance measurement. Cell density in the 3D construct was shown to be proportional to the impedance magnitude of the entire construct. Therefore, perfusion 3D cell culture was performed for up to 5 days and cell proliferation can be monitored by the impedimetric analysis. Moreover, real-time impedimetric monitoring of cell viability under the perfusion of anti-cancer drug in different concentrations was conducted and the impedance magnitude was directly correlated with the cell viability. From the confirmation of the endpoint cell viability assays, a concentration-dependent effect was shown; however, the response of cell viability during the drug treatment was able to be traced by the impedance measurement. The experimental results showed that cell proliferation and chemosensitivity in 3D cell culture format can be monitored by impedance measurement. This microfluidic chip has a high potential to develop a powerful analytical platform for cancer research.

Topics: Antineoplastic Agents; Biosensing Techniques; Cell Culture Techniques; Cell Line, Tumor; Cell Proliferation; Cell Survival; Drug Screening Assays, Antitumor; Electric Impedance; Equipment Design; Humans; Microfluidic Analytical Techniques; Mouth Neoplasms; Sepharose; Tissue Scaffolds

The expression of anchorage independence in malignant oral epithelial cells retrieved from colonies formed in agarose and tumours formed in athymic mice was examined. The original epithelial cell lines were derived from lingual and palatal squamous cell carcinomas induced in rats by the carcinogen 4-nitroquinoline N-oxide. The capacity to express anchorage independence varied considerably between the original cell lines and essentially increased with passage in culture. In three out of four colony-derived subpopulations, the colony-forming efficiency was significantly greater than that of the original cell lines. Xenograft subpopulations expressed higher colony-forming efficiencies than their original counterparts in only two of five cell lines. Undifferentiated tumour xenografts resulted in more homogeneous tumour-derived subpopulations, in contrast to the more heterogeneous cell lines from well-differentiated tumours. The findings demonstrate functional diversity within and between malignant rat oral epithelial cell lines and their colony- and xenograft-derived subpopulations.

Topics: Animals; Carcinoma, Squamous Cell; Culture Media; Fibroblasts; In Vitro Techniques; Keratinocytes; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Mouth Neoplasms; Neoplasm Transplantation; Rats; Rats, Inbred Strains; Sepharose; Tumor Cells, Cultured; Tumor Stem Cell Assay