电穿孔
转染
核转染
菲科尔
细胞培养
化学
巨噬细胞
细胞生物学
分子生物学
生物
生物化学
基因
体外
遗传学
外周血单个核细胞
作者
Neeraja P. Alamuru-Yellapragada,Bandish Kapadia,Kishore V. L. Parsa
标识
DOI:10.1016/j.bbrc.2017.04.043
摘要
Electroporation is the most widely employed method of gene transfer into macrophages which are hard to transfect. RAW 264.7 is a widely used cell line for studying macrophage responses. Electroporation of RAW 264.7 cells with commercial reagents although very efficient is expensive necessitating the development of cost effective alternatives. In this study, we have formulated an economical electroporation buffer for electroporation of RAW 264.7 cells compatible with commercial nucleofector apparatus. We observed that supplementation of membrane fusogenic agents such as Ficoll, PEG and membrane resealing agent, poloxamer P188, enhanced the transfection efficiency of macrophages to a level comparable to the commercially available solutions thereby providing us a cost effective solution for genetic manipulation of macrophages especially in large numbers.
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