UHasselt, BIOMED, Diepenbeek, Belgium; Laboratory of Neuronal Differentiation, VIB Center for the Biology of Disease, Leuven and Center for Human Genetics, KU Leuven, Leuven, Belgium.
UHasselt, BIOMED, Diepenbeek, Belgium.
J Neurosci Methods. 2018 Jan 1;293:169-173. doi: 10.1016/j.jneumeth.2017.09.017. Epub 2017 Sep 29.
Microglia, the resident phagocytic cells of the brain, have recently been the subject of intense investigation given their role in pathology and normal brain physiology. In general, phagocytic cells are hard to transfect with plasmid DNA. The BV2 cell line is a murine cell line of microglial origin which is often used to study this cell type in vitro. Unfortunately, this microglial cell line is, like other phagocytic cells, resistant to transfection.
Magnetofection is a well-established transfection method that combines DNA with magnetic particles which, under the influence of a magnetic field, ensures a high concentration of particles in proximity of cultured cells. Only recently, Glial-Mag was specifically developed for efficient transfection of microglia and microglial cell lines.
Magnetofection with Glial-Mag yielded a transfection efficiency of 34.95% in BV2 cells, 24h after transfection with an eGFP-expressing plasmid. Efficient gene delivery caused a modest and short-lived cell activation (as measured by IL6 secretion) that ceased by 24h after transfection.
Here we show that Glial-Mag magnetofection of BV2 cells yielded a significantly higher transfection efficiency (34.95%) compared to other chemical transfection methods including calcium-phoshate precipication (0.34%), X-tremeGENE (3.30%) and Lipofectamine 2000 (12.51%).
Transfection of BV2 cells using Glial-Mag magnetofection is superior compared to other chemical transfection methods and could be considered as the method of choice to chemically transfect microglial cell lines.
小胶质细胞是大脑中的固有吞噬细胞,由于其在病理学和正常大脑生理学中的作用,最近成为了研究的热点。一般来说,吞噬细胞很难用质粒 DNA 转染。BV2 细胞系是一种源自小胶质细胞的鼠源细胞系,常用于体外研究这种细胞类型。不幸的是,这种小胶质细胞系和其他吞噬细胞一样,对转染有抗性。
磁转染是一种成熟的转染方法,它将 DNA 与磁性颗粒结合,在磁场的影响下,确保磁性颗粒在培养细胞的附近高度集中。最近,Glial-Mag 专门开发用于有效地转染小胶质细胞和小胶质细胞系。
用 Glial-Mag 进行磁转染,在转染含有 eGFP 表达质粒 24 小时后,BV2 细胞的转染效率为 34.95%。有效的基因传递导致适度且短暂的细胞激活(通过 IL6 分泌来衡量),在转染后 24 小时停止。
在这里,我们表明,与其他化学转染方法(包括磷酸钙沉淀(0.34%)、X-tremeGENE(3.30%)和 Lipofectamine 2000(12.51%)相比,Glial-Mag 磁转染 BV2 细胞的转染效率(34.95%)显著更高。
与其他化学转染方法相比,使用 Glial-Mag 磁转染 BV2 细胞系的转染效果更好,可以考虑作为化学转染小胶质细胞系的首选方法。