The invention belongs to the technical field of biological medicines, and particularly relates to amniotic mesenchymal stem cells for treating gonitis as well as a preparation method and application of the amniotic mesenchymal stem cells. The preparation method of the amniotic mesenchymal stem cells comprises the following specific steps: (1) separating to obtain amniotic tissues; (2) digesting, filtering and centrifuging the amniotic membrane tissues to obtain
cell precipitates; (3) precipitating and resuspending the cells, inoculating the cells to a culture medium for culture, digesting, centrifuging, and discarding supernatant to obtain P0-generation amniotic mesenchymal stem cells; and (4) inoculating the P0-generation amniotic mesenchymal stem cells into a culture medium for subculture to obtain the amniotic mesenchymal stem cells. By adding
teprenone and
schaftoside into the basic culture medium, the
cell dryness of the amniotic mesenchymal stem cells can be maintained, and the osteogenic differentiation capability of the amniotic mesenchymal stem cells is improved, so that the gonitis is effectively treated. Meanwhile, fetal calf serum is not added into the basic culture medium, so that the safety of clinical application is improved.