title: A simple way to reconstruct a human 3-d hypodermis: a useful tool for pharmacological functionality.
abstract: BACKGROUND
  Adipose tissue engineering has been hampered by the inability to culture mature adipocytes. Adipose-derived stem cell (ASC) culture opens the way for the preparation of human 3-D hypodermis in large quantities. These models play a role in obesity-related active molecules and slimming agent screening. Moreover, they contribute to a better understanding of the mechanisms underpinning obesity.
  
  
  METHODS
  Freshly extracted ASC from fat tissue were characterized by flow cytometry for CD73, CD90, CD105, HLA-ABC, CD14 and CD45 markers and by Western blot for pref-1. Their differentiation in mature adipocytes was followed by lipid and adiponectin secretion or by oil red O staining and radioimmunoassay. Neosynthesized extracellular matrix (ECM) of 3-D hypodermis was investigated by immunohistochemistry (collagen type I, V and VI) and transmission electron microscopy.
  
  
  RESULTS
  Our results demonstrate that the culture of preadipocytes in proliferation medium for 15 days followed by 16 days of culture in differentiation medium allowed production of the thickest single-layer hypodermis in which preadipocytes and mature adipocytes coexist and synthesize adiponectin and ECM components. Functionality of our 3-D single-layer hypodermis was demonstrated both by a 3.5-fold glycerol production after its stimulation with norepinephrine (adrenergic agonist) and by its slimming after caffeine treatment versus the nontreated 3-D hypodermis.
  
  
  CONCLUSIONS
  This economic 3-D model, easy to prepare and giving reproducible results after the treatment of actives, is useful for pharmacotoxicological trials as an alternative to animal experimentation.
authors: Banque de Tissus et Cellules, Hôpital Edouard Herriot, Lyon, France. charlotte.lequeux @ chu-lyon.fr
authors: C
authors: C
authors: Lequeux
authors: C
authors: C
authors: Auxenfans
authors: A
authors: A
authors: Thépot
authors: A
authors: A
authors: Géloën
authors: V
authors: V
authors: André
authors: O
authors: O
authors: Damour
authors: A
authors: A
authors: Mojallal
mesh_terms: D052242:Adiponectin / Q000378:metabolism; D000273:Adipose Tissue / Q000166:cytology*; D015703:Antigens, CD / Q000378:metabolism; D002110:Caffeine / Q000494:pharmacology; D002454:Cell Differentiation; D002478:Cells, Cultured; D006801:Humans; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D010641:Phenotype; D013234:Stem Cells / Q000166:cytology; D040521:Subcutaneous Tissue / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D052242:Adiponectin; D015703:Antigens, CD; D002110:Caffeine