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
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
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DOI：
10.1210/en.2008-0525
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发表时间：
2008-10-01
期刊：
影响因子：
4.8
通讯作者：
Toda, Shuji
中科院分区：
文献类型：
--
作者：
Sonoda, Emiko;Aoki, Shigehisa;Toda, Shuji
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（MSC），但尚未建立用于分析组织内细胞类型的培养系统。在这里，我们表明，大鼠皮下脂肪组织碎片的三维胶原凝胶培养保持了长期可行的成熟脂肪细胞，产生不成熟的脂肪细胞和MSC样细胞的碎片，使用免疫组化，ELISA和真实的时间RT-PCR。检测成熟脂肪细胞的溴脱氧尿苷摄取。脂联素和瘦素以及脂联素、瘦素和PPAR-gamma的脂肪细胞特异性基因在培养组装中被检测到，而脂肪生成因子胰岛素（20 mU/ml）和炎症相关因子TNF-α（2 nM）分别增加和减少，所有这些都显示。两种梭形细胞类型与油红O-阳性脂滴和MSC标志物（CD 105和CD 44）的表达周围的碎片发展。这些数据表明，脂肪组织器官型培养保留了成熟脂肪细胞的单室结构、增殖能力和一些功能，产生了未成熟脂肪细胞和CD 105（+）/CD 44（+）MSC样细胞。这表明我们的方法将为研究脂肪组织中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。
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.