The invention provides a method of producing pearls on the basis of pearl shell mantel cell culture. The method includes: cutting mantel of a pearl shell into small slices, scattering to obtain single cells by enzymatic depolymerization, performing primary culture prior to subculturing, centrifuging, preparing solution by obtained cells, quantitatively injecting connective tissue in pearl shell mantel with the solution, placing the pearl shells in a culture area, and culturing for one cycle to obtain pearls.