title: Comparative Analysis of Media and Supplements on Initiation and Expansion of Adipose-Derived Stem Cells.
abstract: Adipose-derived stem cells (ASCs) are being tested in clinical trials related to cell-based regenerative therapies. Although most of the current expansion protocols for ASCs use fetal calf serum (FCS), xenogeneic-free medium supplements are greatly desired. This study aims to compare the effect of FCS, human platelet lysate (hPL), and a fully defined medium on the initiation and maintenance of ASC cultures. ASCs obtained from five donors were cultured in five different media: StemPro, Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% hPL, or α-minimum essential medium (A-MEM) supplemented with 5% hPL, 10% hPL, or 10% FCS. The effect of media on proliferation, colony-forming units (CFUs), attachment, and morphology was assessed along with cell size, granularity, and immunophenotype. StemPro greatly compromised the initiation of ASC cultures, which could not survive more than a few passages. Cells cultured in A-MEM proliferated at a faster rate than in DMEM, and hPL significantly enhanced cell size, granularity, and proliferation compared with FCS. All media except StemPro supported CFUs equally well. Analysis of surface markers revealed higher levels of CD73 and CD105 in FCS-cultured ASCs, whereas increased levels of CD146 were found in hPL-cultured cells. Multiparametric flow cytometric analysis performed after seven passages revealed the existence of four distinct ASC subpopulations, all positive for CD73, CD90, and CD105, which mainly differed by their expression of CD146 and CD271. Analysis of the different subpopulations might represent an important biological measure when assessing different medium formulations for a particular clinical application.
authors.affiliation: Laboratory for Stem Cell Research, Department for Health Science and Technology, Aalborg University, Aalborg, Denmark.
authors.forename: Simone
authors.initials: S
authors.lastname: Riis
authors.affiliation: Laboratory for Stem Cell Research, Department for Health Science and Technology, Aalborg University, Aalborg, Denmark.
authors.forename: Frederik Mølgaard
authors.initials: FM
authors.lastname: Nielsen
authors.affiliation: Laboratory for Stem Cell Research, Department for Health Science and Technology, Aalborg University, Aalborg, Denmark.
authors.forename: Cristian Pablo
authors.initials: CP
authors.lastname: Pennisi
authors.affiliation: Laboratory for Stem Cell Research, Department for Health Science and Technology, Aalborg University, Aalborg, Denmark.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Zachar
authors.affiliation: Laboratory for Stem Cell Research, Department for Health Science and Technology, Aalborg University, Aalborg, Denmark trinef@hst.aau.dk.
authors.forename: Trine
authors.initials: T
authors.lastname: Fink
mesh_terms: D017667:Adipocytes / Q000166:cytology; D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D001792:Blood Platelets / Q000166:cytology; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D064987:Cell- and Tissue-Based Therapy; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D044968:Regenerative Medicine; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects
chemical_list: D016895:Culture Media, Serum-Free
keywords: Adipose-derived stem cells; Cell proliferation; Cell therapy; Culture media; Phenotype; Xenogeneic free
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