title: Tissue engineering of white adipose tissue using hyaluronic acid-based scaffolds. I: in vitro differentiation of human adipocyte precursor cells on scaffolds.
abstract: OBJECTIVE
  Reconstruction of soft tissue defects is a challenge in plastic surgery and there is clinical need for adequate solutions. Aim of this study was to develop a biohybrid construct consisting of hyaluronic acid-based scaffolds and human adipocyte precursor cells as a soft tissue filler.
  
  
  METHODS
  Human adipocyte precursor cells were obtained by collagenase digestion of adipose tissue samples and seeded on hyaluronic acid-based spongy scaffolds of various degrees of esterification and pore size using different techniques. After cell attachment, adipose differentiation was induced by defined adipogenic factors under serum-free culture conditions.
  
  
  RESULTS
  Among the five different scaffold types under investigation the highest cell attachment rate was observed for the HYAFF scaffold with 100% esterification and a mean pore size of 400microm (HYAFF 11lp). For inoculation of human adipocyte precursor cells on hyaluronic acid-based scaffolds a "drop-on" technique and low-pressure centrifugation using a Speed Vac airfuge were compared. With respect to efficacy, cell distribution and simpleness the drop-on method proved to be the method of choice. In a serum-free medium supplemented with 66nM insulin, 100nM cortisol and 1microg/ml troglitazone a substantial proportion of cells underwent adipose differentiation as assessed by lipid accumulation and emergence of glycerol-3-phosphate dehydrogenase activity, a lipogenic marker enzyme.
  
  
  CONCLUSIONS
  Hyaluronic acid-based scaffolds appear to be a suitable three-dimensional carrier for the culture and in vitro differentiation of human adipocyte precursor cells.
authors.affiliation: German Diabetes Research Institute, Auf'm Hennekamp 65, 40225 Düsseldorf, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Halbleib
authors.forename: Thomas
authors.initials: T
authors.lastname: Skurk
authors.forename: Claudio
authors.initials: C
authors.lastname: de Luca
authors.forename: Dennis
authors.initials: D
authors.lastname: von Heimburg
authors.forename: Hans
authors.initials: H
authors.lastname: Hauner
mesh_terms: D017667:Adipocytes / Q000166:cytology* / Q000502:physiology*; D000273:Adipose Tissue / Q000166:cytology* / Q000254:growth & development* / Q000378:metabolism; D000293:Adolescent; D000328:Adult; D002448:Cell Adhesion / Q000502:physiology; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods*; D005109:Extracellular Matrix / Q000502:physiology; D005260:Female; D005993:Glycerolphosphate Dehydrogenase / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid; D050356:Lipid Metabolism; D008567:Membranes, Artificial; D008875:Middle Aged; D013234:Stem Cells / Q000166:cytology / Q000502:physiology; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D008567:Membranes, Artificial; D006820:Hyaluronic Acid; D005993:Glycerolphosphate Dehydrogenase