![]() ![]() For example, in the prostate, it has been suggested that terminally differentiated prostate luminal cells give rise to prostate cancer ( 1), as most prostate cancer cells express luminal cell markers but lack basal cell markers ( 2). The exact cellular origin of CSCs needs to be fully elucidated. CSCs represent potential targets for future anticancer therapies, as their capability to serve as reservoirs of cells that cause tumor recurrence has been demonstrated in various types of cancers including prostate and colon cancers. The presence of cancer stem cells (CSCs) in tumors can be a source of such heterogeneity because they have the ability to divide asymmetrically and generate more differentiated cancer clones. It is known that many tumors are composed of heterogeneous populations of cancer cells with different phenotype and characteristics. © 2013 International Society for Advancement of Cytometry Our results revealed that the novel ACCA is a straightforward approach for determining the clonogenic capacity of cancer stem-like cells identified in both cell lines and patient samples. We applied the ACCA to analyze the cloning capacity of different subpopulations of prostate and colon cancer cells based on the expression of the characteristic markers of stem (CD44 and CD133) and cancer stem cells (TROP-2, CD49f, and CD44). The obtained results were also compared with the results of the limiting dilution assay for different cell lines. We tested the new automatic cell-cloning assay (ACCA) on selected cancer cell lines and compared it with the manual approach. Cell plating onto a microplate using an automatic cell deposition unit was performed in a single-cell or dilution rank mode by the fluorescence-activated cell sorting method. ![]() ![]() In this study, we implemented a faster and more precise method for assessing the cloning efficiency of cancer stem-like cells that were characterized and separated using a high-speed cell sorter. However, this approach can also characterize cells with different phenotypes and biological properties, such as stem cells or cancer stem cells. It can be used to determine the cytotoxic effects of various treatments including chemotherapeutics and ionizing radiation. The clonogenic assay is a well-established in vitro method for testing the survival and proliferative capability of cells. ![]()
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