Stem Cell Reports Repor t Identification of Specific Cell-Surface Markers of Adipose-Derived Stem Cells from Subcutaneous and Visceral Fat Depots Wee Kiat Ong, 1 Chuen Seng Tan, 2 Kai Li Chan, 1 Grace Gandi Goesantoso, 1 Xin Hui Derryn Chan, 1 Edmund Chan, 3 Jocelyn Yin, 3 Chia Rou Yeo, 3 Chin Meng Khoo, 3 Jimmy Bok Yan So, 4 Asim Shabbir, 4 Sue-Anne Toh, 3 Weiping Han, 5,6 and Shigeki Sugii 1,5, * 1 Fat Metabolism and Stem Cell Group, Laboratory of Metabolic Medicine, Singapore Bioimaging Consortium, A*STAR, Helios, 11 Biopolis Way, Singapore 138667, Singapore 2 Saw Swee Hock School of Public Health, National University of Singapore, MD3, 16 Medical Drive, Singapore 117597, Singapore 3 Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, MD6, 14 Medical Drive, Singapore 117599, Singapore 4 Department of Surgery, National University Hospital, 5 Lower Kent Ridge Road, Singapore 119074, Singapore 5 Cardiovascular and Metabolic Disorders Program, Duke-NUS Graduate Medical School, 8 College Road, Singapore 169857, Singapore 6 Laboratory of Metabolic Medicine, Singapore Bioimaging Consortium and Metabolism in Human Diseases Unit, Institute of Molecular and Cell Biology, A*STAR, Singapore 138667, Singapore *Correspondence: shigeki_sugii@sbic.a-star.edu.sg http://dx.doi.org/10.1016/j.stemcr.2014.01.002 This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License, which permits non-commercial use, distribution, and reproduction in any medium, provided the original author and source are credited. SUMMARY Adipose-derived stem/stromal cells (ASCs) from the anatomically distinct subcutaneous and visceral depots of white adipose tissue (WAT) differ in their inherent properties. However, little is known about the molecular identity and definitive markers of ASCs from these depots. In this study, ASCs from subcutaneous fat (SC-ASCs) and visceral fat (VS-ASCs) of omental region were isolated and studied. High-content image screening of over 240 cell-surface markers identified several potential depot-specific markers of ASCs. Subsequent studies revealed consistent predominant expression of CD10 in SC-ASCs and CD200 in VS-ASCs across 12 human subjects and in mice. CD10-high-expressing cells sorted from SC-ASCs differentiated better than their CD10-low-expressing counterparts, whereas CD200-low VS-ASCs differentiated better than CD200-high VS-ASCs. The expression of CD10 and CD200 is thus depot-dependent and associates with adipogenic capacities. These markers will offer a valuable tool for tracking and screening of depot-specific stem cell populations. INTRODUCTION White adipose tissue (WAT) has been increasingly appreci- ated as an alternative source of mesenchymal stem cells (MSCs) traditionally isolated from the bone marrow. Subcu- taneous WAT can be isolated by minimally invasive liposuction procedure. Additionally, adipose-derived stem/ stromal cells (ASCs) are relatively abundant in the WAT where as much as 1% of human adipose cells are ASCs as compared to only 0.001%–0.002% MSCs in the bone marrow (Fraser et al., 2006). The differentiation capacity, immunobiological properties, and secretome of ASCs offer tremendous therapeutic potential in regenerative medicine (Ong and Sugii, 2013). By the convention of the International Society for Cellular Therapy (ISCT), MSCs from various sources, including ASC, are defined as being (1) plastic-adherent in the standard cell-culture condition; (2) multipotent, i.e., able to differentiate into osteoblasts, adipocytes, and chondrocytes in vitro; and (3) positive for CD73, CD90, and CD105 and negative for CD11b or CD14, CD19 or CD79a, CD34, CD45, and HLA-DR in their cell-surface immunophenotype (Dominici et al., 2006). In addition, the recent revised statement of ISCT and International Federation for Adipose Therapeutics and Science (IFATS) suggests additional markers for ASCs, which are positive for CD36 and negative for CD106, compared to bone- marrow MSCs (Bourin et al., 2013). Thus, it is beginning to be recognized that MSCs from different origins may have different cell-surface marker expression, but few studies have analyzed their expression differences in a comprehensive manner. Increasing evidence suggests that ASCs derived from WAT of different depot origins are distinct populations of cells that differ in their inherent properties (Macotela et al., 2012; Tchkonia et al., 2005, 2006). A notable func- tional difference is that subcutaneous fat ASCs (SC-ASCs) proliferate at a higher rate and differentiate better than visceral fat ASCs (VS-ASCs) in response to in vitro adipo- genic stimuli (Macotela et al., 2012; Tchkonia et al., 2005). The functional difference of SC- and VS-ASCs together with regional variation in cellular interaction, cir- culation, innervations, and anatomic constraints in the SC and VS depots of WAT are thought to be the underlying fac- tors contributing to pathophysiological variation of these two WAT depots in relation to metabolic homeostasis (Tchkonia et al., 2007, 2013). The SC depot physiologically stores excess lipids thus preventing their deposition into Stem Cell Reports j Vol. 2 j 171–179 j February 11, 2014 j ª2014 The Authors 171