Enrichment isolation of adipose-derived stem/stromal cells from the liquid portion of liposuction aspirates with the use of an adherent column.
OBJECTIVE
Adipose-derived stem/progenitor cells (ASCs) are typically obtained from the lipoaspirates; however, a smaller number of ASCs can be isolated without enzymatic digestion from the infranatant liposuction aspirate fluid (LAF). We evaluated the effectiveness of an adherent column, currently used to isolate mesenchymal stromal cells from bone marrow, to isolate LAF cells.


METHODS
We applied peripheral blood (PB), PB mixed with cultured ASCs (PB-ASC), and LAF solution to the column and divided it into two fractions, the adherent (positive) and the non-adherent (negative) fractions. We compared this method with hypotonic hemolysis (lysis) for the red blood cell count, nucleated cells count and cell compositions as well as functional properties of isolated mesenchymal cells.


RESULTS
The column effectively removed red blood cells, though the removal efficiency was slightly inferior to hemolysis. After column processing of PB-ASC, 60.5% of ASCs (53.2% by lysis) were selectively collected in the positive fraction, and the negative fraction contained almost no ASCs. After processing of LAF solution, nucleated cell yields were comparable between the column and hemolysis; however, subsequent adherent culture indicated that a higher average ASC yield was obtained from the column-positive samples than from the lysis samples, suggesting that the column method may be superior to hemolysis for obtaining viable ASCs. Mesenchymal differentiation and network formation assays showed no statistical differences in ASC functions between the lysis and column-positive samples.


CONCLUSIONS
Our results suggest that a column with non-woven rayon and polyethylene fabrics is useful for isolating stromal vascular fraction cells from LAF solutions for clinical applications.
Department of Plastic Surgery, University of Tokyo School of Medicine, Tokyo, Japan.
Kentaro
K
Doi
Department of Plastic Surgery, University of Tokyo School of Medicine, Tokyo, Japan.
Shinichiro
S
Kuno
Kaneka Corporation, Osaka, Japan.
Akira
A
Kobayashi
Kaneka Corporation, Osaka, Japan.
Takahisa
T
Hamabuchi
Department of Plastic Surgery, University of Tokyo School of Medicine, Tokyo, Japan.
Harunosuke
H
Kato
Department of Plastic Surgery, University of Tokyo School of Medicine, Tokyo, Japan.
Kahori
K
Kinoshita
Department of Plastic Surgery, University of Tokyo School of Medicine, Tokyo, Japan.
Hitomi
H
Eto
Department of Plastic Surgery, University of Tokyo School of Medicine, Tokyo, Japan.
Noriyuki
N
Aoi
Department of Plastic Surgery, University of Tokyo School of Medicine, Tokyo, Japan. Electronic address: kotaro-yoshimura@umin.ac.jp.
Kotaro
K
Yoshimura
D000273:Adipose Tissue / Q000166:cytology*; D002448:Cell Adhesion; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002482:Cellulose; D005434:Flow Cytometry; D006461:Hemolysis; D006801:Humans; D015187:Lipectomy; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D020959:Polyethylene; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
D020959:Polyethylene; D002482:Cellulose; C012024:rayon, purified
adherent column; adipose-derived stem cells; cell isolation; hemolysis; lipoaspirates; liposuction aspirate fluid