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First published online December 1, 2005
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2005-0234v1
24/2/376    most recent
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Submitted on May 24, 2005
Accepted on August 20, 2005

Original Article

The immunophenotype of human adipose derived cells: Temporal changes in stromal- and stem cell-associated markers

James B. Mitchell 1, Kevin McIntosh 1, Sanjin Zvonic 2, Sara Garrett 1, Z. Elizabeth Floyd 2, Amy Kloster 3, Yuan Di Halvorsen 4, Robert W. Storms 5, Brian Goh 2, Gail Kilroy 2, Xiying Wu 2, Jeffrey M. Gimble 2*

1 Cognate Therapeutics, Inc., Baltimore, Maryland
2 Pennington Biomedical Research Center, Baton Rouge, Louisiana
3 Artecel Sciences, Durham, North Carolina
4 CuraGen Corp., Branford, Connecticut
5 Duke University, Durham, North Carolina

* To whom correspondence should be addressed. E-mail: gimblejm{at}pbrc.edu.


  Abstract

Adipose tissue represents an abundant and accessible source of multipotent adult stem cells and is used by many investigators for tissue engineering applications; however, not all laboratories use cells at equivalent stages of isolation and passage. We have compared the immunophenotype of freshly isolated human adipose tissue-derived Stromal Vascular Fraction cells (SVFs) relative to serial passaged Adipose-derived Stem Cells (ASCs). The initial SVFs contained Colony Forming Unit-Fibroblasts (CFU-F) at a frequency of 1:30. Colony Forming Unit-Adipocytes (CFU-Ad) and -Osteoblasts (CFU-Ob) were present in the SVF at comparable frequencies (1:40 and 1:12, respectively). The immunophenotype of the adipose derived cells based on flow cytometry changed progressively with adherence and passage. Stromal cell associated markers (CD13, CD29, CD44, CD63, CD73, CD90, CD166) were initially low on SVFs and increased significantly with successive passages. The stem cell associated marker CD34 was at peak levels in the SVFs and/or early passage ASCs and remained present, although at reduced levels, throughout the culture period. Aldehyde dehydrogenase (ALDH) and the multidrug resistance transport protein (ABCG2), both of which have been used to identify and characterize hematopoietic stem cell, are expressed by SVFs and ASCs at detectable levels. Endothelial cell associated markers (CD31, CD144 or VE-cadherin, VEGF receptor 2, von Willebrand factor) were expressed on SVFs and did not change significantly with serial passage. Thus, the adherence to plastic and subsequent expansion of human adipose-derived cells in fetal bovine serum supplemented medium selects for a relatively homogeneous cell population, enriching for cells expressing a "stromal" immunophenotype, as compared to the heterogeneity of the crude stromal vascular fraction.




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