Tissue engineering of white adipose tissue using hyaluronic acid-based scaffolds. I: in vitro differentiation of human adipocyte precursor cells on scaffolds.
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.
Halbleib
Melanie
M
German Diabetes Research Institute, Auf'm Hennekamp 65, 40225 Düsseldorf, Germany.
Skurk
Thomas
T
de Luca
Claudio
C
von Heimburg
Dennis
D
Hauner
Hans
H
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*
D008567:Membranes, Artificial; D006820:Hyaluronic Acid; D005993:Glycerolphosphate Dehydrogenase