Black M A, Bégin-Heick N
Department of Biochemistry, University of Ottawa, Faculty of Medicine, Canada.
J Cell Biochem. 1995 Aug;58(4):455-63. doi: 10.1002/jcb.240580408.
Stromal vascular cells from epididymal fat pads of lean and obese mice were cultured in a medium (alpha-MEM) containing fetal bovine serum (FBS) and cell replication followed for 11 days. In both types of cells, confluence occurred at 4-5 days, after which virtual growth arrest occurred in lean-mouse cells while replication continued, albeit at a slower rate in obese-mouse cells. Little or no lipid accumulation or glycerol-3-phosphate dehydrogenase (GPDH) activity was observed under these conditions. When a differentiation mixture consisting of insulin, corticosterone and isobutylmethylxanthine was added to the serum-containing alpha-MEM, a proportion of the lean-mouse cells accumulated triglycerides and GPDH activity increased significantly, indicating differentiation. By contrast, little or no differentiation occurred in obese-mouse cells. When cells grown in serum-containing alpha-MEM were transferred to a serum-free defined medium at confluence, extensive differentiation and maturation occurred in lean-mouse cells but not in obese-mouse cells. Similar experiments were conducted in cells isolated from the retroperitoneal fat pad. Although the growth pattern was similar to that of epididymal preadipocytes, the retroperitoneal lean- and obese-mouse cells differentiated more readily than epididymal cells, as shown by the GPDH specific activity. These data suggest that cells from obese mice are resistant to differentiation under conditions that support extensive differentiation in lean-mouse cells.
将来自瘦小鼠和肥胖小鼠附睾脂肪垫的基质血管细胞在含有胎牛血清（FBS）的培养基（α-MEM）中培养，并追踪细胞复制情况达11天。在这两种类型的细胞中，汇合在4 - 5天出现，此后瘦小鼠细胞出现实质生长停滞，而肥胖小鼠细胞的复制继续，尽管速率较慢。在这些条件下几乎未观察到脂质积累或甘油-3-磷酸脱氢酶（GPDH）活性。当将由胰岛素、皮质酮和异丁基甲基黄嘌呤组成的分化混合物添加到含血清的α-MEM中时，一部分瘦小鼠细胞积累了甘油三酯且GPDH活性显著增加，表明发生了分化。相比之下，肥胖小鼠细胞几乎没有发生分化。当在含血清的α-MEM中生长的细胞在汇合时转移到无血清限定培养基中时，瘦小鼠细胞发生了广泛的分化和成熟，而肥胖小鼠细胞则没有。对从腹膜后脂肪垫分离的细胞进行了类似实验。尽管生长模式与附睾前脂肪细胞相似，但如GPDH比活性所示，腹膜后瘦小鼠和肥胖小鼠细胞比附睾细胞更容易分化。这些数据表明，在支持瘦小鼠细胞广泛分化的条件下，肥胖小鼠的细胞对分化具有抗性。
