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Purification of Four Distinct Cell Populations in the Stromal Vascular Fraction of Human Adipose Tissue: Adipogenic Potential and Implications for Soft Tissue Engineering.
Han Li, Ludovic Zimmerlin1,4, Kacey G.Marra, Vera S. Donnenberg2, Albert D Donnenberg, J. Peter Rubin2
Plastic Surgery Research Lab
2010-04-05
Presenter: Han Li
Affidavit:
Director Name:
Author Category: Resident/Fellow
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction
Introduction
Adipose stem cells (ASCs) represent a truly heterogenous population. The aim of this study was to define four distinct populations within the stromal vascular fraction based on surface marker expression, and evaluate the ability of each cell type to differentiate to mature adipocytes.
Materials & Methods
Subcutaneous adipose tissue was obtained by abdominoplasty from human patients. To distinguish isolated cell populations, we performed an 8-color analysis based on the expression of CD3, CD31, CD34, CD45, CD90, CD117 and CD146. Cell proliferation was determined by DNA quantification (Cyquant), and adipocyte differentiation was assessed after exposed to adipogenic culture media.
Results & Discussion
Four cell populations were purified and studied. Candidate perivascular cells (pericytes), defined as CD146+ CD90dim andCD31-CD34-. Two CD31+ endothelial populations were discriminated by CD34 expression and tentatively designated mature endothelial (CD 31+/CD34), and immature endothelial (CD 31+/CD 31-). The CD31-/CD34+ fraction (preadipocyte candidate) was also CD90+, but lacked CD146 expression.
Proliferation was greatest in the CD31-/CD34+ group and slowest in the CD146+ CD90dim group. PPAR gamma and FABP4 mRNA expression was significantly higher in the CD31-/CD34+ group compared with all other populations after exposure to adipgenic medium. The highest proportion of positive Oil Red-O staining was noted in this group, as well.
Conclusions
We have isolated four distinct stromal populations from human adult adipose tissue, the CD31-/CD34+ group is the most prevalent and has the greatest potential for adipogenic differentiation. This cell type appears to hold the most promise for engineering of adipose tissue for reconstructive applications.
