Purification of human adipose-derived stem cells from fat tissues using PLGA/silk screen hybrid membranes

Abstract

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.

MicroRNA-214 Affects Fibroblast Differentiation of Adipose-Derived Mesenchymal Stem Cells by Targeting Mitofusin-2 during Pelvic Floor Dysfunction in SD Rats with Birth Trauma

“…Consistent with these results, many previous studies have also illustrated these kinds of functions of ADMSCs [22, 23]. In addition, it has been shown that bone marrow mesenchymal stem cell (BMSC) transplantation could enhance the closing pressure and leak point pressure in a rat model of SUI [24].…”

Purification and differentiation of human adipose-derived stem cells by membrane filtration and membrane migration methods

“…Human adipose tissue was collected from the fat pads of human patients from the omentum and near the intestines (45–85 years old, six people) after obtaining informed consent. The adipose tissue cell solution (SVF, stromal vascular fraction) was made using a previously reported method162336. The number of hADSCs in the adipose tissue cell solution was evaluated using flow cytometry with antibodies to CD90, CD73, CD40, and CD34 and appropriate isotype antibodies.…”

“…There are several possible methods of isolating hADSCs from SVF solution other than the conventional cell culture method on TCPS dishes: (a) MACS (magnetic-activated cell sorting) method20, (b) FACS (fluorescence-activated cell sorting) method2122, membrane filtration method152324, and membrane migration method16. MACS and FACS use antibodies to purify specific target cells such as CD166, CD105, CD90, CD73, CD44, and CD29 expressing cells (CD166 + , CD105 + , CD90 + , CD73 + , CD44 + , and CD29 + cells).…”

“…Therefore, the membrane filtration method and/or migration method could be an attractive method for the isolation of hADSCs from SVF solution because of short processing time, sterility, and simplicity of the method. In particular, hADSCs can be isolated from SVF solution within 30 min using the membrane filtration method, whereas the conventional culture method typically takes 1–2 weeks to purify hADSCs from SVF solution152324.…”

“…hADSCs were also purified by the membrane filtration method using different type of porous membranes. hADSCs were isolated from SVF solution in permeation solution by filtration through porous silk screen/poly(lactide-co-glycolic acid) membranes using the sieving effect23.…”