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Over the last 10 years, huge advances have been made worldwide in the adult stem cell field. Several donor sites can be used for harvesting mesenchymal stem cells (MSC), bone marrow and adipose tissue being the most frequent. The latter is easily harvested by liposuction and, in most patients, a large quantity of MSC can be obtained without harm to the donor (Casteilla et al., 2004). In 2001, Zuk and co-workers showed that a human lipoaspirate contains multipotent cells and may represent an alternative stem cell source to bone marrow-derived MSC. Adipose-derived MSC are capable of proliferation in monolayer culture and multilineage differentiation in response to inductive conditions, and thus have potential clinical application (Bailey et al., 2010; Fraser et al., 2008; Rigotti et al., 2009;). However, research and clinical groups have distinct protocols to isolate and manipulate these cells, differing in the type and concentration of the enzyme used, time and conditions of incubation for adipose tissue digestion and methods of cell culture. These methodological differences result in diverse characteristics of the cells isolated and varied functional results. Therefore, the development of a standardized and reproducible method of isolating MSC as well as standard techniques for functional characterization is fundamental to validate cells for its use in therapeutic protocols. A temptative for functional characterization of MSC was recently proposed by the International Society for Cellular Therapy (ISCT) (Dominici et al., 2006). According to this statement, MSC must: (i) be plastic-adherent in standard cultures; (ii) express a mesenchymal set of surface molecules; (iii) differentiate into osteoblasts, adipocytes and chondroblasts in vitro. The detection of these mesenchymal set of surface molecules as well as the differentiation assays can be performed on adipose-tissue MSC population after their isolation in laboratory and will be carefully described in this chapter.

Processing of Lipoaspirate Samples for Optimal Mesenchymal Stem Cells Isolation