title: Isolation, Expansion, and Immortalization of Human Adipose-Derived Mesenchymal Stromal Cells from Biopsies and Liposuction Specimens.
abstract: Human adipose tissue has proven to be an abundant, accessible, and rich source of adult mesenchymal stromal cells, suitable for tissue engineering and regenerative medicine. However, a major complication in fully investigating these cells may derive from their limited life span.Although methods to isolate, expand, and immortalize these cells have been widely reported in the literature, exhaustive explanations on the problems that can be encountered during these processes and how these can be solved have never been described. It is of fundamental importance to follow a common protocol to achieve reliable and reproducible results. Here, we describe a protocol to isolate and expand human adipose stromal cells from specimens obtained from tissue biopsies and liposuction surgical interventions. Finally, we broadly describe the cell immortalization technique, and particular attention is paid to some of the apparently "secondary" aspects.
authors.affiliation: Medestea Research and Production Laboratories, Consorzio CARSO, Strada Pro.le Valenzano Casamassima, Km 3, Bari, Valenzano, 70010, Italy. luigi.balducci.lb@gmail.com.
authors.forename: Luigi
authors.initials: L
authors.lastname: Balducci
authors.affiliation: Department of Cerebrovascular Diseases, Fondazione IRCCS Neurological Institute Carlo Besta, Milan, Italy.
authors.forename: Giulio
authors.initials: G
authors.lastname: Alessandri
mesh_terms: D001706:Biopsy; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D006801:Humans; D016130:Immunophenotyping; D015187:Lipectomy; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D013234:Stem Cells; D050151:Subcutaneous Fat / Q000166:cytology*
keywords: Adipose-derived stromal cells; Enzymatic digestion; Expansion; Fat biopsy; Human adipose tissue; Immortalization; Isolation; Liposuction; Stromal vascular fraction