Establishment of a preadipocyte cell line derived from mature adipocytes of GFP transgenic mice and formation of adipose tissue
Establishment of a preadipocyte cell line derived from mature adipocytes of GFP transgenic mice and formation of adipose tissue
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DOI：
10.1007/s00441-008-0593-9
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发表时间：
2008-06-01
影响因子：
3.6
通讯作者：
Kano, Koichiro
中科院分区：
文献类型：
--
作者：
Nobusue, Hiroyuki;Endo, Tsuyoshi;Kano, Koichiro
我们从绿色荧光蛋白（GFP）转基因小鼠的皮下脂肪组织中获得的成熟脂肪细胞建立了前脂肪细胞系。通过胶原酶消化和过滤从皮下脂肪组织中分离出含有成熟脂肪细胞的漂浮顶层。荧光激活细胞分选和显微镜分析表明，漂浮细胞部分由高度均质的脂肪细胞群组成，不含脂肪基质血管细胞。分离的成熟脂肪细胞去分化为成纤维细胞样细胞，并在天花板培养物中活跃增殖。体外研究表明，这些细胞可以以与 3T3-L1 前脂肪细胞相同的方式再分化为成熟脂肪细胞。在我们的细胞增殖过程中没有观察到分化模式的变化。它们被成功维持并分化了至少 22 代。我们将这些细胞命名为去分化脂肪（DFAT-GFP）细胞。当 DFAT-GFP 细胞皮下植入 C57BL/6N 小鼠时，它们发育出高度血管化的脂肪垫，其形态与正常皮下脂肪组织相似，由 GFP 阳性细胞组成；然而，植入的 3T3-L1 细胞对小鼠没有这样的影响。我们的结论是，DFAT-GFP 细胞提供了一个模型，使我们能够研究体内和体外脂肪细胞分化和脂肪组织形成的机制。
We established a preadipocyte cell line from mature adipocytes obtained from subcutaneous fat tissue of green fluorescent protein (GFP) transgenic mice. The floating top layer, containing mature adipocytes, was isolated from subcutaneous fat tissue by collagenase digestion and filtration. Fluorescence-activated cell sorting and microscopic analysis revealed that the floating cell fraction comprised a highly homogeneous adipocyte population with no adipose stromal-vascular cells. Isolated mature adipocytes dedifferentiated into fibroblast-like cells and actively proliferated in ceiling culture. In vitro studies showed that the cells could redifferentiate into mature adipocytes in an identical way to 3T3-L1 preadipocytes. No changes in the differentiation pattern were observed during the propagation of our cells. They were successfully maintained and differentiated for at least 22 passages. We named these cells dedifferentiated fat (DFAT-GFP) cells. When DFAT-GFP cells were implanted subcutaneously into C57BL/6N mice, they developed highly vascularized fat pads that morphologically resembled normal subcutaneous adipose tissue and consisted of GFP-positive cells; however, implanted 3T3-L1 cells did not have such an effect on the mice. We conclude that DFAT-GFP cells provide a model that should enable us to study the mechanisms of adipocyte differentiation and adipose tissue formation in vivo and in vitro.