小鼠棕色脂肪原代培养模型的建立

目的:探索小鼠棕色脂肪细胞原代培养的方法?方法:取C57BL/6J小鼠棕色脂肪组织,采用胶原酶消化过滤法获得梭形细胞,对培养出来的细胞进行形态学观察,诱导分化后用油红O染色法染色定性,荧光定量PCR检测棕色脂肪标志基因表达情况?结果:培养出的梭形细胞成分均一,增殖旺盛,诱导分化后分化率高,经油红O染色证实为脂肪细胞,荧光定量PCR检测棕色脂肪标志基因UCP-1表达量明显升高?结论:从C57BL/6J小鼠棕色脂肪组织中可以分离出具有很强增殖?分化能力的前棕色脂肪细胞,这种棕色脂肪细胞原代培养模型的建立为在体外进一步研究棕色脂肪的功能提供了良好的基础?

Establishment of culture model for primary mouse brown adipocyte precursor cells

Objective: To establish a culture method for primary mouse brown adipocyte precursor cells. Methods: Fibroblast-like cells were collected from C57BL/6j mice brown adipose tissue. The morphological changes of the cultured cells were observed,the intracytoplasmic lipid of the culture cells was determined using oil red O staining,and the expression of the brown adipocyte specific gene was determined by real time PCR. Results: The cultured fibroblast-like cells showed highly homogeneous appearance with active proliferation and differentiate into mature adipocytes. Oil red O staining,morphological observation,and expression of brown adipocyte specific gene UCP-1 verified these cells as brown adipocyte. Conclusion: Brown adipocyte precursor cells are present in C57BL/6j mice brown adipose tissue and possess the potential to proliferate and differentiate into mature brown adipocyte. The establishment of primary culture of mouse brown adipocyte precursor cells is important for furth studies of the function of brown adipocyte in vitro.