title: Isolation of adipose-derived stem cells: a comparison among different methods.
abstract: Adipose-derived stromal cells (ASCs) are usually isolated by digestion with collagenase. We have compared alternative methods to isolate ASCs in a more economically viable protocol. Nine protocols using red blood cells lysis buffer solution, trypsin, collagenase and centrifugation were compared; the isolation rate, cell viability, expansion rate, immunophenotype and differentiation in adipogenic and osteogenic lineages were analyzed. ASCs were isolated and successfully maintained by digestion with trypsin. Cells presented similar immunophenotypes, adipogenic differentiation and in vitro proliferation but an osteogenic differentiation capacity up to seven times higher than ASCs isolated by collagenase. This alternative protocol is thus efficient and more cost-effective than the commonly-used methods and may represent a promising protocol for obtaining ASCs for bone tissue engineering.
authors: Complexo Hospitalar Santa Casa de Porto Alegre, Rua Annes Dias 295, Porto Alegre, RS, 90020-090, Brazil.
authors: Carolina Franke
authors: CF
authors: Markarian
authors: Gianna Zaffari
authors: GZ
authors: Frey
authors: Maiele Dornelles
authors: MD
authors: Silveira
authors: Eduardo Mainieri
authors: EM
authors: Chem
authors: Adriana Rosa
authors: AR
authors: Milani
authors: Pedro Bins
authors: PB
authors: Ely
authors: Ana Paula
authors: AP
authors: Horn
authors: Nance Beyer
authors: NB
authors: Nardi
authors: Melissa
authors: M
authors: Camassola
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D016130:Immunophenotyping; D013048:Specimen Handling / Q000379:methods; D013234:Stem Cells / Q000502:physiology*