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Discrete adipose-derived stem cell subpopulations may display differential functionality after in vitro expansion despite convergence to a common phenotype distribution.

Abstract：

BACKGROUND:Complex immunophenotypic repertoires defining discrete adipose-derived stem cell (ASC) subpopulations may hold a key toward identifying predictors of clinical utility. To this end, we sorted out of the freshly established ASCs four subpopulations (SPs) according to a specific pattern of co-expression of six surface markers, the CD34, CD73, CD90, CD105, CD146, and CD271, using polychromatic flow cytometry. METHOD:Using flow cytometry-associated cell sorting and analysis, gating parameters were set to select for a CD73+CD90+CD105+ phenotype plus one of the four following combinations, CD34-CD146-CD271- (SP1), CD34-CD146+CD271- (SP2), CD34+CD146+CD271- (SP3), and CD34-CD146+CD271+ (SP4). The SPs were expanded 700- to 1000-fold, and their surface repertoire, trilineage differentiation, and clonogenic potential, and the capacity to support wound healing were assayed. RESULTS:Upon culturing, the co-expression of major epitopes, the CD73, CD90, and CD105 was maintained, while regarding the minor markers, all SPs reverted to resemble the pre-sorted population with CD34-CD146-CD271- and CD34-CD146+CD271- representing the most prevalent combinations, followed by less frequent CD34+CD146-CD271- and CD34+CD146+CD271- variants. There was no difference in the efficiency of adipo-, osteo-, or chondrogenesis by cytochemistry and real-time RT-PCR or the CFU capacity between the individual SPs, however, the SP2CD73+90+105+34-146+271- outperformed others in terms of wound healing. CONCLUSIONS:Our study shows that ASCs upon culturing inherently maintain a stable distribution of immunophenotype variants, which may potentially disguise specific functional properties of particular downstream lines. Furthermore, the outlined approach suggests a paradigm whereby discrete subpopulations could be identified to provide for a therapeutically most relevant cell product.

journal_name

Stem Cell Res Ther

journal_title

Stem cell research & therapy

authors

Nielsen FM,Riis SE,Andersen JI,Lesage R,Fink T,Pennisi CP,Zachar V

doi

10.1186/s13287-016-0435-8

subject

Has Abstract

pub_date

2016-12-01 00:00:00

pages

177

issue

issn

1757-6512

pii

10.1186/s13287-016-0435-8

journal_volume

pub_type

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Discrete adipose-derived stem cell subpopulations may display differential functionality after in vitro expansion despite convergence to a common phenotype distribution.