农业生物技术学报2013，Vol.21Issue(4)：379-387,9.DOI:10.3969/j.issn.1674-7968.2013.04.001
基于一种新的天花板培养方法分析猪成熟脂肪细胞去分化过程中关键基因表达模式
Analysis of Expression Patterns of Key Genes During the Pig(Sus scrofa)Mature Adipocyte Dedifferentiation Based on a Novel Ceiling Culture Model
摘要
Abstract
When mature adipocytes are subjected to an in vitro dedifferentiation strategy referred to as ceiling culture, these mature adipocytes can revert to a more primitive phenotype and gain cell proliferative ability. We refer to these cells as dedifferentiated fat (DFAT) cells. Because of those cells possessing multilineage differentiation potential, dedifferentiation has become a significant research topic in recent years. Here we firstly isolated pig mature adipocytes and indentified them by GENMED staining.Then, in order to clarify the mechanism of mature adipocyte dedifferentiation, a novel ceiling culture model was developed and the mRNA expression of marker genes during dedifferentiation couse was analysized using Real-time PCR. Finally, we assessed the effect of 1 μmol/L rosiglitazone on mature adipocyte dedifferentiation. The results showed that 98.7% of isolated cells were mononuclear mature adipocytes and these adipocytes could efficiently dedifferentiate into DFAT cells under a novel ceiling culture model. During the course of dedifferentiation, the mRNA levels of adipogenic marker peroxisome proliferator activated receptor y (PPARy), adipocyte- type fatty acid- binding protein (aP2) and lipoprotein lipase (LPL) were up- regulated by 8, 3 and 7.5 folds, respectively. While the lipolytic marker hormone- sensitive lipase (HSL) and Adipose triglyceride lipase (ATGL) were increased about 40 and 10 folds in mRNA level, respectively. In addition, the dedifferentiation was remarkably suppressed by treatment with 1 μmol/mL rosiglitazone. These results indicate that it is mainly a lipolytic course companying with little ability of adipogenesis that mature adipocytes dedifferentiate into DFAT cells, and this will provide new theoretical reference for future research.关键词
猪成熟脂肪细胞/新式天花板培养法/去分化/脂解Key words
Pig mature adipocyte/ A novel ceiling culture/ Dedifferentiation/ Lipolysis引用本文复制引用
宋子仪,史新娥,杨浩,高倩,赵丽丽,杨公社..基于一种新的天花板培养方法分析猪成熟脂肪细胞去分化过程中关键基因表达模式[J].农业生物技术学报,2013,21(4):379-387,9.基金项目
国家转基因生物新品种培育重大专项(No.2011ZX08006-005)和国家973计划项目(No.2012CB124705) （No.2011ZX08006-005）
