Purification of human adipose-derived stem cells from fat tissues using PLGA/silk screen hybrid membranes
Purification of human adipose-derived stem cells from fat tissues using PLGA/silk screen hybrid membranes
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DOI：
10.1016/j.biomaterials.2014.02.004
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发表时间：
2014-05-01
期刊：
影响因子：
14
通讯作者：
Higuchi, Akon
中科院分区：
文献类型：
--
作者：
Chen, Da-Chung;Chen, Li-Yu;Higuchi, Akon
研究了通过聚（丙交酯-共-乙醇酸）/丝网杂化膜使用膜过滤从人脂肪组织细胞（基质血管组分）纯化人脂肪源性干细胞（hADSC）。膜过滤方法在再生医学中是有吸引力的，因为它们减少了纯化hADSC所需的时间（即，少于30分钟），而常规培养方法需要5-12天。将表达间充质干细胞标志物CD 44、CD 73和CD 90的hADSC浓缩在来自杂交膜的渗透溶液中。来自混合膜的渗透溶液中的细胞上的表面标志物CD 44、CD 73和CD 99的表达在统计学上显著高于原代脂肪组织细胞的表达，所述混合膜使用含有50 × 10（4）个细胞的18 mL进料溶液获得，这表明可以通过膜过滤方法在渗透溶液中纯化hADSC。在渗透液中可以成功分离表达干细胞相关标志物CD 34的细胞，而CD 34（+）细胞不能通过常规培养方法纯化。与来自人脂肪组织悬浮液的细胞相比，基于其碱性磷酸酶活性、其osterix基因表达以及通过茜素红S和von Kossa染色的矿化分析结果，渗透溶液中的hADSC表现出上级的成骨分化能力。这些结果表明，能够成骨分化的hADSC优先渗透通过杂化膜。(C)2014爱思唯尔有限公司版权所有。
The purification of human adipose-derived stem cells (hADSCs) from human adipose tissue cells (stromal vascular fraction) was investigated using membrane filtration through poly(lactide-co-glycolic acid)/silk screen hybrid membranes. Membrane filtration methods are attractive in regenerative medicine because they reduce the time required to purify hADSCs (i.e., less than 30 min) compared with conventional culture methods, which require 5-12 days. hADSCs expressing the mesenchymal stem cell markers CD44, CD73, and CD90 were concentrated in the permeation solution from the hybrid membranes. Expression of the surface markers CD44, CD73, and CD99 on the cells in the permeation solution from the hybrid membranes, which were obtained using 18 mL of feed solution containing 50 x 10(4) cells, was statistically significantly higher than that of the primary adipose tissue cells, indicating that the hADSCs can be purified in the permeation solution by the membrane filtration method. Cells expressing the stem cell-associated marker CD34 could be successfully isolated in the permeation solution, whereas CD34(+) cells could not be purified by the conventional culture method. The hADSCs in the permeation solution demonstrated a superior capacity for osteogenic differentiation based on their alkali phosphatase activity, their osterix gene expression, and the results of mineralization analysis by Alizarin Red S and von Kossa staining compared with the cells from the suspension of human adipose tissue. These results suggest that the hADSCs capable of osteogenic differentiation preferentially permeate through the hybrid membranes. (C) 2014 Elsevier Ltd. All rights reserved.