A new organotypic culture of adipose tissue fragments maintains viable mature adipocytes for a long term, together with development of immature adipocytes and mesenchymal stem cell-like cells.
Adipose tissue that consists of mature and immature adipocytes is suggested to contain mesenchymal stem cells (MSCs), but a culture system for analyzing their cell types within the tissue has not been established. Here we show that three-dimensional collagen gel culture of rat sc adipose tissue fragments maintained viable mature adipocytes for a long term, producing immature adipocytes and MSC-like cells from the fragments, using immunohistochemistry, ELISA, and real time RT-PCR. Bromodeoxyuridine uptake of mature adipocytes was detected. Adiponectin and leptin, and adipocyte-specific genes of adiponectin, leptin, and PPAR-gamma were detected in culture assembly, whereas the lipogenesis factor insulin (20 mU/ml) and inflammation-related agent TNF-alpha (2 nm) increased and decreased, respectively, all of their displays. Both spindle-shaped cell types with oil red O-positive lipid droplets and those with expression of MSC markers (CD105 and CD44) developed around the fragments. The data indicate that adipose tissue-organotypic culture retains unilocular structure, proliferative ability, and some functions of mature adipocytes, generating both immature adipocytes and CD105+/CD44+ MSC-like cells. This suggests that our method will open up a new way for studying both multiple cell types within adipose tissue and the cell-based mechanisms of obesity and metabolic syndrome.
Department of Pathology and Biodefense, Faculty of Medicine, Saga University, Nabeshima 5-1-1, Saga 849-8501, Japan.
Emiko
E
Sonoda
Shigehisa
S
Aoki
Kazuyoshi
K
Uchihashi
Hidenobu
H
Soejima
Sachiko
S
Kanaji
Kenji
K
Izuhara
Seiji
S
Satoh
Noboru
N
Fujitani
Hajime
H
Sugihara
Shuji
S
Toda
D017667:Adipocytes / Q000166:cytology*; D054392:Adipokines / Q000235:genetics; D000273:Adipose Tissue / Q000166:cytology* / Q000502:physiology*; D000818:Animals; D002455:Cell Division / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D003094:Collagen; D003470:Culture Media / Q000494:pharmacology; D000071063:Endoglin; D005455:Fluorescent Antibody Technique; D005782:Gels; D015870:Gene Expression / Q000502:physiology; D018960:Hyaluronan Receptors / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D020738:Leptin / Q000235:genetics; D008055:Lipids; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D009924:Organ Culture Techniques / Q000379:methods*; D051381:Rats; D017208:Rats, Wistar
D054392:Adipokines; D003470:Culture Media; D000071063:Endoglin; C501740:Eng protein, rat; D005782:Gels; D018960:Hyaluronan Receptors; D047908:Intracellular Signaling Peptides and Proteins; D020738:Leptin; D008055:Lipids; D003094:Collagen