title: Decellularized adipose tissue microcarriers as a dynamic culture platform for human adipose-derived stem/stromal cell expansion.
abstract: With the goal of designing a clinically-relevant expansion strategy for human adipose-derived stem/stromal cells (ASCs), methods were developed to synthesize porous microcarriers derived purely from human decellularized adipose tissue (DAT). An electrospraying approach was applied to generate spherical DAT microcarriers with an average diameter of 428 ± 41 μm, which were soft, compliant, and stable in long-term culture without chemical crosslinking. Human ASCs demonstrated enhanced proliferation on the DAT microcarriers relative to commercially-sourced Cultispher-S microcarriers within a spinner culture system over 1 month. ASC immunophenotype was maintained post expansion, with a trend for reduced expression of the cell adhesion receptors CD73, CD105, and CD29 under dynamic conditions. Upregulation of the early lineage-specific genes PPARγ, LPL, and COMP was observed in the ASCs expanded on the DAT microcarriers, but the cells retained their multilineage differentiation capacity. Comparison of adipogenic and osteogenic differentiation in 2-D cultures prepared with ASCs pre-expanded on the DAT microcarriers or Cultispher-S microcarriers revealed similar adipogenic and enhanced osteogenic marker expression in the DAT microcarrier group, which had undergone a higher population fold change. Further, histological staining results suggested a more homogeneous differentiation response in the ASCs expanded on the DAT microcarriers as compared to either Cultispher-S microcarriers or tissue culture polystyrene. A pilot chondrogenesis study revealed higher levels of chondrogenic gene and protein expression in the ASCs expanded on the DAT microcarriers relative to all other groups, including the baseline controls. Overall, this study demonstrates the promise of applying dynamic culture with tissue-specific DAT microcarriers as a means of deriving regenerative cell populations.
authors.affiliation: Department of Chemical Engineering, Queen's University, 19 Division St., Kingston, ON, K7L 3N6, Canada; Human Mobility Research Center, Kingston General Hospital, 76 Stuart St., Kingston, ON, K7L 2V7, Canada.
authors.forename: Claire
authors.initials: C
authors.lastname: Yu
authors.affiliation: Biomedical Engineering Graduate Program, Claudette MacKay Lassonde Pavilion, The University of Western Ontario, London, ON, N6A 5B9, Canada.
authors.forename: Anna
authors.initials: A
authors.lastname: Kornmuller
authors.affiliation: Department of Anatomy and Cell Biology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, N6A 5C1, Canada.
authors.forename: Cody
authors.initials: C
authors.lastname: Brown
authors.affiliation: Department of Chemical Engineering, McMaster University, 1280 Main Street West, Hamilton, ON, L8S 4L7, Canada.
authors.forename: Todd
authors.initials: T
authors.lastname: Hoare
authors.affiliation: Department of Anatomy and Cell Biology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, N6A 5C1, Canada; Department of Chemical and Biochemical Engineering, Thompson Engineering Building, The University of Western Ontario, London, ON, N6A 5B9, Canada. Electronic address: lauren.flynn@uwo.ca.
authors.forename: Lauren E
authors.initials: LE
authors.lastname: Flynn
mesh_terms: D017667:Adipocytes / Q000166:cytology* / Q000502:physiology; D000273:Adipose Tissue / Q000737:chemistry*; D061249:Batch Cell Culture Techniques / Q000379:methods*; D002214:Capsules / Q000737:chemistry*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D049109:Cell Proliferation / Q000502:physiology; D002474:Cell-Free System; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000502:physiology; D006801:Humans; D008904:Miniaturization; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D002214:Capsules
keywords: Adipose-derived stem/stromal cells (ASCs); Dynamic culture; Extracellular matrix (ECM); Microcarriers; Microenvironment; Stem cell expansion