IADR Abstract Archives
Isolation and charaterisation of oral cancer stem cells
Objectives: Cancer stem cells (CSCs) are a subset of the tumour cell population which possess the capability for indefinite self-renewal, are highly tumorigenic, and are also resistant to anticancer therapy. Identifying and isolating CSCs is crucial to study their characteristics with the aim of specifically targeting and eradicating them.
We hypothesize that CSCs can be functionally isolated from oral cancer cell lines and used two different assays (cell adhesion and chemo-resistance) to investigate the stem cell characteristics of these cells, compared to unsorted cells.
Methods: Cancer stem-like cells were isolated from two oral cancer cell lines (H357 and SCC4) on the basis of their rapid adherence (within 10 minutes) to fibronectin (FN) or their resistance to conventional chemotherapy (5μM cisplatin for 3 days). The stem cell characteristics of the isolated cells were investigated using FACs analysis, proliferation and colony forming assays.
Results: Cells isolated using both methods expressed significantly increased levels of stem cell markers CD24, CD44 and CD29 compared to unsorted cells. In addition, the growth rate of early adherent cells was two-fold lower and the colony forming efficiency three-fold higher than unsorted cells. Moreover, these cells exhibited increased cisplatin resistance compared to unsorted cells. Conversely, the cisplatin-resistant cells adhered to fibronectin more highly than unsorted cells and exhibited a higher growth rate compared to unsorted cells after a second cisplatin exposure.
Conclusions: Rapid adherence to FN and chemo-resistance effectively isolates a sub-population of cells from cell lines which show stem cell-like characteristics. The increased chemoresistance of early adherent cells and the increased adherence of cisplatin-resistant cells suggest that these two methods are isolating the same population of CSCs. Future work will investigate the stability in culture of these CSCs with the aim of developing strategies to eradicate them from cultures and subsequently tumours in vivo
We hypothesize that CSCs can be functionally isolated from oral cancer cell lines and used two different assays (cell adhesion and chemo-resistance) to investigate the stem cell characteristics of these cells, compared to unsorted cells.
Methods: Cancer stem-like cells were isolated from two oral cancer cell lines (H357 and SCC4) on the basis of their rapid adherence (within 10 minutes) to fibronectin (FN) or their resistance to conventional chemotherapy (5μM cisplatin for 3 days). The stem cell characteristics of the isolated cells were investigated using FACs analysis, proliferation and colony forming assays.
Results: Cells isolated using both methods expressed significantly increased levels of stem cell markers CD24, CD44 and CD29 compared to unsorted cells. In addition, the growth rate of early adherent cells was two-fold lower and the colony forming efficiency three-fold higher than unsorted cells. Moreover, these cells exhibited increased cisplatin resistance compared to unsorted cells. Conversely, the cisplatin-resistant cells adhered to fibronectin more highly than unsorted cells and exhibited a higher growth rate compared to unsorted cells after a second cisplatin exposure.
Conclusions: Rapid adherence to FN and chemo-resistance effectively isolates a sub-population of cells from cell lines which show stem cell-like characteristics. The increased chemoresistance of early adherent cells and the increased adherence of cisplatin-resistant cells suggest that these two methods are isolating the same population of CSCs. Future work will investigate the stability in culture of these CSCs with the aim of developing strategies to eradicate them from cultures and subsequently tumours in vivo