目的:探讨细胞传代培养对胰岛素样生长因子Ⅱ受体(insulin-like growth factor Ⅱ receptor,Igf2r)和Mest基因表达的影响,以及它们在不同部位来源的成纤维样细胞(fibroblast-like cells)中的表达。方法:利用实时定量PCR法检测印迹基因Igf2r和Mest在来源于鼠耳和尾尖的成纤维样细胞中的表达。结果:在鼠尾成纤维样细胞的传代培养中,随着培养代数的增加,Igf2r的表达减弱,Mest的表达无明显变化;Igf2r在鼠尾成纤维样细胞中的表达高于鼠耳。结论:传代培养影响Igf2r基因的表达,Igf2r在不同部位来源的成纤维样细胞中的表达具有差异,为供核细胞的选择及后续的基因印迹研究奠定了基础。
Objective: To investigate the effects of cell subcultivation on the expression and significance of insulin-like growth factor Ⅱ receptor (Igf2r) and Mest gene, as well as their expressions in fibroblast-like cells derived from different positions. Methods: Real time PCR was employed to detect the expressions of imprinting gene Igf2r and Mest in fibroblast-like cells derived from the ear and tail tip of mice. Results: In the subcuhivation of fibroblast-like cells derived from the tail of mice, the Igf2r expression was decreased along with the increase of different passages, and no significant difference of the Mest expression was observed. The Igf2r expression in the fibroblast-like cells derived from the tail tip was higher than that derived from the ear of mice. Conclusion: Serial subcuhivation has effect on Igf2r expression, and the Igf2r expression in fibroblast-like ceils derived from different positions is different, which provides a basis for the selection of donor cells and further study on genetic imprinting.