The invention relates to a method for preparing cells overexpressing exogenous genes through electrotransfection, which comprises the following steps: treating the cells with a
cell electrotransfection pretreatment
reagent before electrotransfection, and the
cell electrotransfection pretreatment
reagent contains
cholesterol derivatives. The invention also relates to the exogenous
gene overexpressed
cell prepared by the method, application of the cell electrotransfection pretreatment
reagent in preparation of the exogenous
gene overexpressed cell, and a cell electrotransfection kit. The cell electrotransfection kit comprises a cell electrotransfection medium component and a cell treatment component. The cell treatment component comprises a
cholesterol derivative. The method provided by the invention can greatly improve the
survival rate of cells after electrotransfection and the proportion and quantity of cells with positive exogenous
gene expression.