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Chen D.-C.,National Central University | Chen L.-Y.,National Central University | Ling Q.-D.,National Central University | Ling Q.-D.,Cathay Medical Research Institute | And 12 more authors.
Biomaterials | Year: 2014

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 × 104 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. © 2014 Elsevier Ltd. Source


Higuchi A.,National Central University | Higuchi A.,King Saud University | Ling Q.-D.,National Central University | Ling Q.-D.,Cathay Medical Research Institute | And 8 more authors.
Laboratory Investigation | Year: 2015

Induced pluripotent stem cells (iPSCs) provide a platform to obtain patient-specific cells for use as a cell source in regenerative medicine. Although iPSCs do not have the ethical concerns of embryonic stem cells, iPSCs have not been widely used in clinical applications, as they are generated by gene transduction. Recently, iPSCs have been generated without the use of genetic material. For example, protein-induced PSCs and chemically induced PSCs have been generated by the use of small and large (protein) molecules. Several epigenetic characteristics are important for cell differentiation; therefore, several small-molecule inhibitors of epigenetic-modifying enzymes, such as DNA methyltransferases, histone deacetylases, histone methyltransferases, and histone demethylases, are potential candidates for the reprogramming of somatic cells into iPSCs. In this review, we discuss what types of small chemical or large (protein) molecules could be used to replace the viral transduction of genes and/or genetic reprogramming to obtain human iPSCs. © 2015 USCAP, Inc All rights reserved. Source


Wang P.-Y.,National Central University | Lee H.H.-C.,Cathay General Hospital | Lee H.H.-C.,National Central University | Higuchi A.,National Central University | And 15 more authors.
Journal of Materials Chemistry B | Year: 2015

The stem cell fates of pluripotency and differentiation are regulated by not only soluble biological cues but also insoluble biochemical cues (i.e., extracellular matrix (ECM)) and the physical cues of cell culture biomaterials (i.e., elasticity). We investigated the maintenance of pluripotency and the differentiation lineages of human amniotic fluid-derived stem cells (hAFSCs) cultured on poly(vinyl alcohol-co-itaconic acid) (PVA) hydrogels grafted with several types of ECM and corresponding oligopeptides in expansion medium. hAFSCs cultured on soft PVA hydrogels (12.2 kPa) that were grafted with oligopeptides derived from fibronectin and vitronectin showed high pluripotency, which was evaluated by Oct4, Sox2 and Nanog expression. The hAFSCs grown on soft PVA hydrogels (12.2 kPa) grafted with each oligopeptide showed higher pluripotency, as assessed by Oct4 and Nanog expression, than hAFSCs grown on stiff PVA hydrogels (25.3 kPa) grafted with the same oligopeptides and a much higher pluripotency than those grown on rigid tissue-culture polystyrene dishes. Soft biomaterials appeared to be adequate to maintain the pluripotency of hAFSCs. Surprisingly, hAFSCs that showed higher pluripotency on PVA hydrogels grafted with oligopeptides derived from fibronectin and vitronectin also expressed higher levels of early differentiation markers for three germ layers in expansion medium. This result suggests that hAFSCs are heterogeneous and that this population contains highly pluripotent stem cells and stem cells that can be easily differentiated. © 2015 The Royal Society of Chemistry. Source


Higuchi A.,National Central University | Higuchi A.,National Health Research Institute | Higuchi A.,Cathay Medical Research Institute | Chuang C.-W.,National Central University | And 10 more authors.
Journal of Membrane Science | Year: 2011

Adipose-derived stem cells (ADSCs) were purified from mice adipose-tissue cell solutions by the conventional culture method and the membrane filtration (i.e., batch-type filtration and perfusion-type filtration) method. The ADSCs expressing the mesenchymal stem cell marker CD73 were concentrated in a recovery solution through one sheet of polyurethane (PU) foaming membranes with a pore size of 11 μm, and in a permeate solution through five sheets of Nylon mesh filters with a pore size of 11 μm, by the perfusion-type filtration method. This provided a concentration of cells expressing the marker that was 1.7 times higher than that of cells in the primary adipose-tissue cell solution. The ADSCs in the recovery solution that went through the PU foaming membranes but not through the Nylon mesh filters showed greater adipogenic and osteogenic differentiation ability than the cells contained in the primary adipose-tissue cell solution. The perfusion-type filtration effectively recovered ADSCs with a greater ability to differentiate into adipocytes and osteoblasts than the cells recovered by batch-type filtration. These results suggested that the ADSCs with adipogenic and osteogenic differentiation ability tended to adhere to PU membranes but not to Nylon mesh filters when using perfusion-type filtration. The relationship between the ratio of cells expressing the mesenchymal stem cell surface marker (i.e., CD73) and the adipogenic and osteogenic differentiation ability of the cells was also investigated. © 2010 Elsevier B.V. Source


Wu C.-H.,National Central University | Lee F.-K.,Cathay General Hospital | Suresh Kumar S.,National Central University | Ling Q.-D.,Cathay Medical Research Institute | And 10 more authors.
Biomaterials | Year: 2012

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. © 2012 Elsevier Ltd. Source

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