The invention relates to exfoliated cell preservation solution and a preparation method thereof. The preservation solution comprises the following components in percentage by weight: 0.6 percent of sodium chloride, 0.06 percent of potassium chloride, 2 percent of glucose, 20 percent of ethanol, 3 percent of 1-3 propylene glycol, 2 percent of glycerin, 0.15 percent of glycine, 0.1 percent of mucolytic agent, 0.2 percent of cell protective agent, 0.1 percent of heparin sodium and 0.05N of sodium benzoate-benzoic acid buffer solution. A PH value is adjusted by 1 percent sodium hydroxide or hydrochloric acid solution to ensure that the PH of the preservation solution is 5.8. By the exfoliated cell preservation solution, cells can be well preserved and the preservation time is more than 10 days; cell nucleuses are clear, and cytoplasm is stretched; red blood cells and mucus are effectively removed; the cells are dispersed and flatly laid and an acid phosphatase (ACP) enzyme is well preserved simultaneously; and the exfoliated cell preservation solution is suitable for ACP enzyme staining.