Document Detail


Adipose tissue engineering in three-dimensional levitation tissue culture system based on magnetic nanoparticles.
MedLine Citation:
PMID:  23017116     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
White adipose tissue (WAT) is becoming widely used in regenerative medicine/cell therapy applications, and its physiological and pathological importance is increasingly appreciated. WAT is a complex organ composed of differentiated adipocytes, stromal mesenchymal progenitors known as adipose stromal cells (ASC), as well as endothelial vascular cells and infiltrating leukocytes. Two-dimensional (2D) culture that has been typically used for studying adipose cells does not adequately recapitulate WAT complexity. Improved methods for reconstruction of functional WAT ex vivo are instrumental for understanding of physiological interactions between the composing cell populations. Here, we used a three-dimensional (3D) levitation tissue culture system based on magnetic nanoparticle assembly to model WAT development and growth in organoids termed adipospheres. We show that 3T3-L1 preadipocytes remain viable in spheroids for a long period of time, while in 2D culture, they lose adherence and die after reaching confluence. Upon adipogenesis induction in 3T3-L1 adipospheres, cells efficiently formed large lipid droplets typical of white adipocytes in vivo, while only smaller lipid droplet formation is achievable in 2D. Adiposphere-based coculture of 3T3-L1 preadipocytes with murine endothelial bEND.3 cells led to a vascular-like network assembly concomitantly with lipogenesis in perivascular cells. Adipocyte-depleted stromal vascular fraction (SVF) of mouse WAT cultured in 3D underwent assembly into organoids with vascular-like structures containing luminal endothelial and perivascular stromal cell layers. Adipospheres made from primary WAT cells displayed robust proliferation and complex hierarchical organization reflected by a matricellular gradient incorporating ASC, endothelial cells, and leukocytes, while ASC quickly outgrew other cell types in adherent culture. Upon adipogenesis induction, adipospheres derived from the SVF displayed more efficient lipid droplet accumulation than 2D cultures. This indicates that 3D intercellular signaling better recapitulates WAT organogenesis. Combined, our studies show that adipospheres are appropriate for WAT modeling ex vivo and provide a new platform for functional screens to identify molecules bioactive toward individual adipose cell populations. This 3D methodology could be adopted for WAT transplantation applications and aid approaches to WAT-based cell therapy.
Authors:
Alexes C Daquinag; Glauco R Souza; Mikhail G Kolonin
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2012-11-02
Journal Detail:
Title:  Tissue engineering. Part C, Methods     Volume:  19     ISSN:  1937-3392     ISO Abbreviation:  Tissue Eng Part C Methods     Publication Date:  2013 May 
Date Detail:
Created Date:  2013-03-19     Completed Date:  2014-01-17     Revised Date:  2014-05-08    
Medline Journal Info:
Nlm Unique ID:  101466663     Medline TA:  Tissue Eng Part C Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  336-44     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
3T3-L1 Cells
Adipocytes / cytology
Adipogenesis
Adipose Tissue / blood supply,  physiology*
Adipose Tissue, White
Animals
Cell Differentiation
Cell Proliferation
Coculture Techniques
Magnetic Phenomena*
Mice
Nanoparticles / chemistry*
Spheroids, Cellular / cytology
Tissue Culture Techniques / methods*
Tissue Engineering / methods*
Grant Support
ID/Acronym/Agency:
1R21DK090752/DK/NIDDK NIH HHS; R01 DK088131/DK/NIDDK NIH HHS; R01DK088131/DK/NIDDK NIH HHS
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