Centrifugal countercurrent chromatography to elucidate surface differences of adipose tissue-derived stem cells.
The current methods of isolation of adipose tissue-derived stem cells result in a heterogeneous population that might interfere with their differentiation potential and makes it difficult to compare the results between different groups. Partition in aqueous two-phase systems is one of the few techniques that separate cells on the basis of surface properties, gentle enough to isolate fragile cell types in isotonic conditions without altering their structure, and can be easily scaled. In this study, stem cells isolated from human adipose tissue seeded and expanded in vitro were fractionated by using centrifugal countercurrent distribution in an aqueous two-phase system. The separated subpopulations revealed the high heterogeneity of adipose tissue-derived stem cell samples. Comparative partition analyses showed that aging induces a loss of heterogeneity, which is not due to a loss of cell viability associated to age. The phosphatidylserine externalization, an apoptotic feature, is the main factor in cell partition that results in a decreased hydrophobicity of the cell surface. This procedure may be suitable for separating adipose tissue-derived stem cell populations enriched in some functional and/or structural surface characteristics. The possibility of a very effective separation of different subpopulations in opposite phases would be an interesting development of the method.
Department of Biochemistry and Molecular and Cellular Biology, School of Veterinary Medicine, University of Zaragoza, Zaragoza, Spain.
Ramiro
R
Sebastián
Víctor
V
Calvin
Noelia
N
Mendoza
Rosaura
R
Pérez-Pé
David
D
García
Carmen
C
Carreras
José A
JA
Cebrián-Pérez
Teresa
T
Muiño-Blanco
D000273:Adipose Tissue / Q000737:chemistry* / Q000166:cytology; D002454:Cell Differentiation; D003377:Countercurrent Distribution / Q000379:methods*; D006801:Humans; D013234:Stem Cells / Q000737:chemistry* / Q000166:cytology; D013499:Surface Properties