| dc.description.abstract | Cancer stem cells (CSCs, cancer-initiating cells) typically comprise 1%–5% of the total tumor cell population. This cell population is considered to be primarily responsible for tumor initiation, growth, and metastasis. However, it is hard to distinguish cancer cells from other cells (i.e. fibroblasts, blood cells, etc.) in primary tissue after enzyme digestion process. Therefore, We develop membrane filtration and migration method to target and purify rare primary CSCs based on their high migration mobility characteristics compared with other tissue cells. It is expected to establish the primary colon cancer cell line from primary tumor tissue by the membrane filtration and migration method for the development of patient specific therapy in clinical application.
We designed a membrane filtration method to purify and further establish primary colon cancer cell line from human colon cancer tissue. Nylon mesh filters with pore size of 11 μm and 20 μm and PLGA (poly (lactic-co-glycolic acid))-silk screen membranes with different PLGA concentration 3%, 5%, and 10% as the membranes are used in this study where PLGA has biodegradation and biocompatibility properties. With increasing PLGA concentration, the thickness of the PLGA-silk screen membranes also increased. Nylon has unique physical and chemical properties. These properties can also apply to cell culture process. In the primary cancer cell extraction procedures, the colon cancer tissue was digested by collagenase type IV at first. After digestion of the tissue, the primary colon cancer cell solution was permeated through the membranes with different pore size and different degree of biocompatibility. These membranes were used to capture primary colon cancer cells, and the cells on the membranes were expected to migrate into the culture dishes during the cultivation of the membranes in the culture medium (i.e., membrane migration method) after the filtration of colon cancer tissue solution through the membranes. Afterwards, the expression of cancer stem cell markers, CD133 and CD44 was evaluated by flow cytometry. Furthermore, the malignancy index of primary cells and the cells purified by the membrane filtration and migration method was evaluated by soft agarose colony forming assay. It is expected that the cells migrate from the membranes will have higher expression of the colon cancer stem cell surface markers. The similar results may be obtained in soft agarose colony forming assay where the cells show higher colony forming efficiency and bigger colony size. The goal of this project is to establish a patient-specific colon cancer cell line by the membrane filtration and/or migration method for precision medicine in the future.
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