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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

Departments of Pathology and Biodefense (E.S., S.A., K.U., S.T.), Biomolecular Sciences (H.S., S.K., K.I.), and Surgery (S.S.), Faculty of Medicine, Saga University, Saga 849-8501, Japan; Chiba Institute of Science (N.F.), Chiba 288-0025, Japan; and International University of Health and Welfare (S.H.), The School of Rehabilitation Sciences, Fukuoka 831-8501, Japan

Address all correspondence and requests for reprints to: Miss Emiko Sonoda, Master of Medicine/Dr. Shuji Toda, Department of Pathology and Biodefense, Faculty of Medicine, Saga University, Nabeshima 5-1-1, Saga 849-8501, Japan. E-mail: todas{at}cc.saga-u.ac.jp.

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- were detected in culture assembly, whereas the lipogenesis factor insulin (20 mU/ml) and inflammation-related agent TNF- (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.