Xiao W-L, Shi B, Zheng Q, Wang Y, Huang L, Li S, Lu Y, Wu M
Department of Oral and Maxillofacial Surgery, West China College of Stomatology, Sichuan University, Chengdu, People's Republic of China.
Int J Oral Maxillofac Surg. 2007 May;36(5):429-34. doi: 10.1016/j.ijom.2006.12.008. Epub 2007 Apr 27.
Non-syndromic cleft of the lip and/or palate is one of the most common birth defects in humans. Embryonic palatal mesenchymal (EPM) cells are an attractive source for investigating embryonic palatal development. In this study, we developed a highly efficient transfection method for murine EPM (MEPM) cells. MEPM cells were transfected with the plasmid pEGFP-N1 using two non-viral methods: nucleofection and lipofection. Nucleofection provided a much better rate of gene transfer than lipofection particularly in MEPM cells. The methylenetetrahydrofolate reductase (MTHFR) gene is an important candidate for involvement in the pathogenesis of this birth defect. The RNA interference plasmid of MTHFR was constructed and nucleofected into MEPM cells. Successful transfection resulted in a remarkable reduction in the expression of MTHFR. Taken together, the results indicate that nucleofection is highly efficient for MEPM cell transfection, and that this approach may be useful for investigating gene function in the process of palatogenesis.
非综合征性唇裂和/或腭裂是人类最常见的出生缺陷之一。胚胎腭间充质（EPM）细胞是研究胚胎腭发育的一个有吸引力的细胞来源。在本研究中，我们开发了一种针对小鼠EPM（MEPM）细胞的高效转染方法。使用两种非病毒方法（核转染和脂质体转染）将质粒pEGFP-N1转染到MEPM细胞中。核转染比脂质体转染具有更好的基因转移率，特别是在MEPM细胞中。亚甲基四氢叶酸还原酶（MTHFR）基因是参与这种出生缺陷发病机制的一个重要候选基因。构建了MTHFR的RNA干扰质粒并将其核转染到MEPM细胞中。成功转染导致MTHFR的表达显著降低。综上所述，结果表明核转染对MEPM细胞转染非常高效，并且这种方法可能有助于研究腭发生过程中的基因功能。
