Biomarker Discovery and Development

Colon Cancer (CC) Biomarkers

The Advantages of AlfaGene's System ovER cURRENT mETHODOLOGIES

Previous methods of experimentation have utilized malignant transformed cell lines, isolated intestinal epithelial cells (IECs), and animal models. In all cases, however, inaccuracies have arisen in extrapolating from the data obtained from these methods, since each differs markedly from in vivo stem and epithelial cells and each method has inherent limitations (Table 1).  For example, malignant cell lines usually have chromosomal abnormalities that cause instability. Tumor lines also differ significantly from the in vivo cell type being examined, cell line to cell line, and passage to passage.  Transformed cell lines demonstrate altered growth control due to the transfection of viral DNA, such as the large T-antigen from SV40.  The use of freshly isolated epithelial or stem cells is often complicated, not only by the concurrent isolation and contamination with a mixture of several additional cell types including T cells, B cells, or macrophages (especially when inflamed tissue is used as cell source) and low yields, but also, by the fact that the isolation procedure can alter the phenotype of the cells by cleaving certain molecules from the cell surface. Furthermore, surface and crypt epithelial cells may have different phenotypes and functions and are not usually fully represented in separated populations.  Findings with animal model systems show that these do not always precisely mimic the functional scenarios of human intestinal epithelial cells in vivo.

Table 1. Comparison of previous models of stem and intestinal epithelial cell (IEC) vs. our newly developed stem, human intestinal primary epithelial cell (HIPEC), and  human intestinal tumorogenic epithelial cell (HITEC) model.

AlfaGene’s technology enables the generation and long-term maintenance in culture of stem and nontransformed primary epithelial cell lines derived from tissue samples, thus avoiding the issues associated with changes induced by transformation, limited yields, and non uniformity.