A hybrid-membrane migration method to isolate high-purity adipose-derived stem cells from fat tissues

Abstract

Human adipose-derived stem cells (hADSCs) exhibit heterogeneous characteristics, indicating various genotypes and differentiation abilities. The isolated hADSCs can possess different purity levels and divergent properties depending on the purification methods used. We developed a hybrid-membrane migration method that purifies hADSCs from a fat tissue solution with extremely high purity and pluripotency. A primary fat-tissue solution was permeated through the porous membranes with a pore size from 8 to 25 μm, and the membranes were incubated in cell culture medium for 15-18 days. The hADSCs that migrated from the membranes contained an extremely high percentage (e.g., >98%) of cells positive for mesenchymal stem cell markers and showed almost one order of magnitude higher expression of some pluripotency genes (Oct4, Sox2, Klf4 and Nanog) compared with cells isolated using the conventional culture method.

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

“…Typically, higher expression of MSC surface markers on hADSCs is found with increasing passage number14171819. However, we found that expression of some pluripotent genes such as Oct3/4, Nanog , and Klf4 , which were typical genes used for the transduction of somatic cells in reprogramming into hiPSCs1234, decreased after culture of cells in SVF solution on TCPS in our previous study16, whereas the pluripotent gene expression of the cells in SVF solution could be maintained after purification by filtration via porous membranes16. These surprising results indicate that hADSC characteristics depend on the method used for isolation of hADSCs from SVF solution.…”

“…1a). Mesenchymal stem cell (MSC) marker expression typically increases after SVF solution is cultured on conventional tissue culture polystyrene (TCPS) dishes141516. MSC surface markers in SVF solution often show less than 10–20% expression, whereas MSC surface markers of the cells after culture on TCPS (i.e.…”

“…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).…”

Microfluidic systems for stem cell-based neural tissue engineering

“…Furthermore it was represented that cells obtained from human liposuction fat have special ability to divide into other cell types. These MSCs were called adipose-drived stem cells (ASCs) 55 . ASCs are abundant and can be harvested with minimal invasive procedure.…”

“…HSCs derived from bone marrow (that can generate all types of different blood cells) 137 , and different types of MSCs found in other tissues such as skeletal, bone marrow are some examples of this subject. Adipose derived SCs (ADSCs) are multipotent MSCs that exist in subcutaneous fat deposits and are easily isolated from material obtained using lipoplasty (fat removal) 55 .…”