The invention relates to nucleated pearl lamella conditioning fluid in the technical field of pearl culture and application of the nucleated pearl lamella conditioning fluid. The conditioning fluid comprises the following constituents, by mass-to-volume concentration percentage, 1-5% of acid red, 0.01-5% of chondroitin, 0.1-5% of mafenide, 0.1-5% of taurine, 0.75% of sodium chloride, 0.1-5% of hyriopsis cumingii polysaccharide, and 0.05-3% of vitronectin, and allowance being water. The invention further relates to the application of the nucleated pearl lamella conditioning fluid in the pearl culture. The nucleated pearl lamella conditioning fluid can strengthen cell viability, prevent cell infection, and accelerate cell division and wound healing. When the nucleated pearl lamella conditioning fluid is applied to culture pearls, the death rate of the pearls is obviously reduced, and the rate of remaining nucleuses is obviously improved. Meanwhile, lamellas can be rapidly attached to lamella insertion shells, and the lamella insertion shells can rapidly proliferate to grow into pearl sacs. The nucleated pearl lamella conditioning fluid is simple in method, easy to achieve, wide in application range and remarkable in effect, is used for cultivating nucleated pearls by fresh water pearl mussels, and has wide application prospects.