Isolation and culture of human adipose-derived stem cells from subcutaneous and visceral white adipose tissue compartments

Ge, Xiaojia, Leow, Shi Chi, Sathiakumar, Durgalakshmi, Stünkel, Walter, Shabbir, Asim, So, Jimmy, Lomanto, Davide, and McFarlane, Craig (2016) Isolation and culture of human adipose-derived stem cells from subcutaneous and visceral white adipose tissue compartments. Bio-protocol, 6 (22).

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Human Adipose-derived Stem/Stromal Cells (ASCs) have been widely used in stem cell and obesity research, as well as clinical applications including cell-based therapies, tissue engineering and reconstruction. Compared with mesenchymal stem cells (MSCs) derived from other tissues such as umbilical cord and bone marrow, isolation of ASCs from human white adipose tissue (WAT) has great advantages due to its rich tissue source and simple surgical procedure. In this detailed protocol we describe a protocol to isolate and characterize ASCs from human WAT. Molecular characterization of isolated ASCs was performed through surface marker expression profiling using flow cytometry. Adipogenic capacity of the isolated ASCs was confirmed through inducing adipogenic differentiation and Oil Red O staining of lipid. This protocol provides researchers with the tools to culture and assess purity and adipogenic differentiation capacity of human ASCs, which can then be utilized for required downstream in vitro applications.

This protocol has been modified from Baglioni et al. (2009), Baglioni et al. (2012), and van Harmelen et al. (2005) to describe in detail a complete technique to isolate and subsequently characterize human ASCs from human WAT biopsies. This protocol has been utilized to isolate and characterize human ASCs from both subcutaneous and visceral WAT. The isolated human ASCs show high purity and demonstrate adipogenic differentiation capacity in vitro.

Item ID: 52361
Item Type: Article (Short Note)
ISSN: 2331-8325
Funders: Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research, Singapore (A*STAR)
Date Deposited: 13 Aug 2019 02:59
FoR Codes: 06 BIOLOGICAL SCIENCES > 0601 Biochemistry and Cell Biology > 060103 Cell Development, Proliferation and Death @ 30%
10 TECHNOLOGY > 1004 Medical Biotechnology > 100404 Regenerative Medicine (incl Stem Cells and Tissue Engineering) @ 70%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 100%
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