Sonoda Emiko, Aoki Shigehisa, Uchihashi Kazuyoshi, Soejima Hidenobu, Kanaji Sachiko, Izuhara Kenji, Satoh Seiji, Fujitani Noboru, Sugihara Hajime, Toda Shuji
Department of Pathology and Biodefense, Faculty of Medicine, Saga University, Nabeshima 5-1-1, Saga 849-8501, Japan.
Endocrinology. 2008 Oct;149(10):4794-8. doi: 10.1210/en.2008-0525. Epub 2008 Jun 5.
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.
由成熟和未成熟脂肪细胞组成的脂肪组织被认为含有间充质干细胞（MSCs），但尚未建立用于分析组织内细胞类型的培养系统。在此，我们展示了大鼠皮下脂肪组织片段的三维胶原凝胶培养可长期维持存活的成熟脂肪细胞，利用免疫组织化学、酶联免疫吸附测定和实时逆转录聚合酶链反应从这些片段中产生未成熟脂肪细胞和类间充质干细胞。检测到成熟脂肪细胞的溴脱氧尿苷摄取。在培养物中检测到脂联素和瘦素，以及脂联素、瘦素和过氧化物酶体增殖物激活受体γ的脂肪细胞特异性基因，而脂肪生成因子胰岛素（20 mU/ml）和炎症相关因子肿瘤坏死因子α（2 nM）分别增加和降低了它们所有的表达。在片段周围出现了带有油红O阳性脂滴的纺锤形细胞类型以及表达间充质干细胞标志物（CD105和CD44）的细胞类型。数据表明脂肪组织器官型培养保留了单泡结构、增殖能力以及成熟脂肪细胞的一些功能，产生未成熟脂肪细胞和CD105+/CD44+类间充质干细胞。这表明我们的方法将为研究脂肪组织内的多种细胞类型以及肥胖和代谢综合征的细胞机制开辟一条新途径。
