Department of Pharmaceutical Sciences, Southern Illinois University Edwardsville School of Pharmacy, Edwardsville, IL 62026, USA.
Biotechniques. 2022 Dec;73(6):273-279. doi: 10.2144/btn-2022-0054. Epub 2022 Nov 18.
There are various approaches in which one can isolate microglia from murine brains, such as immunomagnetic, density gradient, FACS and differential adhesive methods. In this procedure a modified flask-tapping approach was used due to its simplicity and reproducibility. Our protocol requires only a single step to isolate the microglia from the mixed cell population. Once the microglia were isolated, we characterized cell purity, microglial morphology and phagocytic activity. The single-step protocol, without the need for additional astrocyte or oligodendrocyte separation, allows microglial cells to be used immediately for experimental purposes. The protocol is low-cost and can be performed in any lab with standard cell-culture equipment.
有多种方法可以从鼠脑中分离小胶质细胞,如免疫磁珠法、密度梯度离心法、流式细胞术和差分式黏附法。在本实验中,由于其简单性和可重复性,我们使用了改良的瓶击法。我们的方案仅需一步即可从小胶质细胞与混合细胞群中分离小胶质细胞。从小胶质细胞中分离出来后,我们对细胞纯度、小胶质细胞形态和吞噬活性进行了表征。该单步方案无需额外分离星形胶质细胞或少突胶质细胞,可立即将小胶质细胞用于实验目的。该方案成本低,任何配备标准细胞培养设备的实验室均可进行。