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The isolation and differentiation of human adipose-derived stem cells using membrane filtration

机译:膜过滤法分离和分化人脂肪干细胞

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Human adipose-derived stem cells (hADSCs) were purified from a suspension of human adipose tissue cells (stromal vascular fraction) by the conventional culture method and by membrane filtration through polyurethane (PU) foam membranes. hADSCs can be obtained from a suspension of human adipose tissue cells using the membrane filtration method in less than 30 min, whereas the conventional culture method requires 5-12 days. hADSCs that express the mesenchymal stem cell markers CD44, CD73, and CD90 were concentrated in the recovery solution from the PU membranes; no hADSCs were isolated in the permeate. After filtration, the cells expressing the mesenchymal stem cell markers were 3-4.5 times more concentrated than in the initial suspension of human adipose tissue cells, with the level of concentration depending on the surface modification of the PU membrane. Cells expressing the stem cell-associated marker CD34 could be successfully isolated in the recovery solutions, whereas CD34 + cells could not be purified by the conventional culture method. The hADSCs in the recovery solution demonstrated a superior capacity for osteogenic differentiation than did the cells in the suspension of human adipose tissue cells. These results suggested that the hADSCs with the capability for osteogenic differentiation adhered to the PU membranes.
机译:通过常规培养方法并通过聚氨酯(PU)泡沫膜进行膜过滤,从人脂肪组织细胞悬液(间质血管部分)中纯化人脂肪来源的干细胞(hADSC)。使用膜过滤法可在不到30分钟的时间内从人脂肪组织细胞悬液中获得hADSC,而常规培养方法则需要5-12天。表达间充质干细胞标志物CD44,CD73和CD90的hADSCs浓缩在PU膜的回收溶液中。在渗透物中未分离出hADSC。过滤后,表达间充质干细胞标记的细胞的浓度是人脂肪组织细胞初始悬浮液的3-4.5倍,其浓度水平取决于PU膜的表面修饰。表达干细胞相关标志物CD34的细胞可以在回收液中成功分离出来,而CD34 +细胞无法通过常规培养方法纯化。与人类脂肪组织细胞悬液中的细胞相比,回收液中的hADSCs具有更好的成骨分化能力。这些结果表明具有成骨分化能力的hADSCs粘附在PU膜上。

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