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目的:对比改进的“天花板”组织块培养法与消化培养法培养前脂肪细胞的优劣。方法:两种方法获得的前脂肪细胞,培养3天,显微镜下检测自分化情况,MTT、流式细胞仪检测其增殖情况和细胞周期。培养9天,绘制生长曲线。使用含胰岛素、甲状腺素T3和转铁蛋白(ITT)的无血清培养基诱导获得的前脂肪细胞5天,拍照测细胞直径,油红染色法检测其分化情况。结果:改进的“天花板”组织块培养法培养的第一代自分化存在,但是发生自分化的细胞很少。增殖和分化结果均无显著性差异。但是此法在同等情况下获得细胞较多,所需经费较少。结论:与消化培养法相比较而言,改进的“天花板”组织块培养法是一种更加简单、经济的获得大鼠前脂肪细胞的方法。
OBJECTIVE: To compare the advantages and disadvantages of the improved “ceiling” tissue culture method and digestion culture method for culturing preadipocytes. Methods: The preadipocytes obtained by the two methods were cultured for 3 days, the autodetection was examined under a microscope, and the proliferation and cell cycle were detected by MTT and flow cytometry. Cultivate for 9 days and draw the growth curve. The preadipocytes were induced using serum-free medium containing insulin, thyroxine T3 and transferrin (ITT) for 5 days. The diameters of the cells were measured and the differentiation was detected by oil red staining. RESULTS: The first generation of self-differentiated cultures cultured with the modified “ceiling” tissue culture method existed, but few cells were self-differentiated. There was no significant difference in proliferation and differentiation. However, this method obtains more cells under the same conditions and requires less funds. Conclusion: Compared with the digestion culture method, the improved “ceiling” tissue culture method is a simpler and economical method to obtain rat preadipocytes.