| Summary: | 碩士 === 國立中央大學 === 化學工程與材料工程研究所 === 100 === Adipose-derived stem cells (ADSCs) are a promising cell source in regenerative medicine, of particular utility for cell therapies and tissue engineering, because adipose tissue can easily be harvested in large quantities compared to bone marrow, and ADSCs have high proliferation rates in culture. ADSCs are isolated from adipose tissue by liposuction and centrifugation followed by cultivation on cell culture dishes for at least one passage. The cultivation of cells derived from adipose tissue is necessary to purify ADSCs (i.e., “the culture method” for the purification of ADSCs) because the adipose tissue contains not only ADSCs but also adipose and other types of cells. The culture process for the purification of ADSCs requires several days, at minimum. If ADSCs can be purified from adipose tissue in a short period of time (i.e., less than 2 hrs) by using a cell purification device such as the membrane filtration method, cell therapy and tissue engineering applications using autologous ADSCs might become more efficient.
Therefore, we investigated the purification of human ADSCs from a digested solution of adipose tissue by the membrane filtration method in this study, and we compared the purity of ADSCs and the differentiation ability of ADSCs into osteoblasts after purification by the membrane filtration method and the conventional cell culture method.
We investigated two filtration methods to purify hADSC, i.e., batch-type and perfusion-type filtration methods. Main differences between these two filtration methods are cell flow direction to the membranes. Polyurethane foaming membranes having 5-12 μm of pore size were used as the membranes for the separation of hADSCs from human adipose tissue. The surface marker of ADSCs (e.g., CD73 and CD90) in the cells in the permeate and recovery solutions were analyzed by flow cytometry whether the mesenchymal stem cells were enriched after permeation through the membranes. The differentiation of cells into osteoblasts, which were separated by the membrane filtration method was evaluated to confirm the enriched hADSC in the permeate solution through the membranes by culture of the cells in induced medium of osteogenic differentiation.
We, further, investigated the isolation of ADSCs by the membrane filtration method through surface-modified PU membranes having with various nanosegments (e.g., -NH2, -SO3H, -OH, and -COOH) and ECMs, and compared the isolation efficiency of ADSCs purified through nonmodified PU membranes and surface-modified PU membranes. We found that the cells separated through PU membranes by the perfusion method showed high popuration of ADSCs from surface marker analysis and the highest osteogenic differentiation ability.
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