Taylor A, Rogers M J, Tosh D, Coxon F P
Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
Calcif Tissue Int. 2007 Feb;80(2):132-6. doi: 10.1007/s00223-006-0245-6. Epub 2007 Feb 3.
Mature osteoclasts and their precursors are notoriously difficult to transfect using nonviral approaches, a limitation that represents a major technical obstacle in the study of osteoclast biology. Here, we describe a simple electroporation method using Amaxa Nucleofector technology that results in efficient transfection of human blood-derived osteoclast precursors, which can be differentiated in subsequent culture to generate mature osteoclasts that retain expression of the transgene. Moreover, since these osteoclasts maintain the ability to resorb dentine, this technique could prove useful for assessing the role of specific genes/proteins in osteoclast function.
众所周知,使用非病毒方法转染成熟破骨细胞及其前体非常困难,这一限制是破骨细胞生物学研究中的一个主要技术障碍。在此,我们描述了一种使用Amaxa Nucleofector技术的简单电穿孔方法,该方法可有效转染源自人血的破骨细胞前体,这些前体在后续培养中可分化生成保留转基因表达的成熟破骨细胞。此外,由于这些破骨细胞保持了吸收牙本质的能力,该技术可能有助于评估特定基因/蛋白质在破骨细胞功能中的作用。