The invention discloses a megalobrama amblycephala bone tissue culture method. The method includes steps: 1), selecting megalobrama amblycephala of 3 months old as an experimental material; 2), taking caudal vertebra, costa and its connective tissue around of megalobrama amblycephala; 3), flushing with PBS, wetting a tissue block in fetal calf serum, and using forceps to put the tissue block at the bottom of a culture bottle; 4), putting the culture bottle into a culture box for inverted culture; 5), daily observing cell growth and replacing a culture medium; 6), performing trypsin digestion passage after cell convergence degree reaches 80% to obtain megalobrama amblycephala primary bone tissue cells; 7), using trypsin for digestion passage of the cells when convergence degree of the megalobrama amblycephala primary bone tissue cells reaches 70-80%, centrifuging before re-suspending the cells, and inoculating the cells into two new culture bottles to continue culture to obtain passage cells. The method is easy to implement and simple and convenient to operate, the problems of limitation on sample individual size and limitation on using of a trypsin digestion method are solved, and a base material is provided for related study on fish skeleton growth and development and the like in the future.