The invention discloses a separation and culture method for germ cells of gonadal tissue of
tegillarca granosa. The separation and culture method comprises the following steps: firstly, sucking gonadal tissue of
tegillarca granosa by a sterile
injector under a sterile condition, carrying out disinfection, performing digestive enzymolysis on the gonadal tissue at 23 DEG C with a
mixed solution containing 0.05% of pancreatin and 0.2% of
collagenase, collecting a
cell suspension, carrying out
centrifugation, removing a supernatant, suspending precipitated cells with a disinfection
buffer solution, controlling the horizontal
centrifugation speed at 2600 rpm / m when taking
sperm cells from male gonadal tissue, controlling the horizontal
centrifugation speed at 1600 rpm / m when taking egg cells from female gonadal tissue, taking the
cell suspension, paving a 6-hole plate, carrying inverted
adherent culture in a biochemical culture box at 26 DEG C for 24 h, then setting the plate right, adding1 mL of culture medium to each hole, then, replacing a half of culture medium every 2 d, carrying out constant-temperature standing culture for 5-10 d, and performing
primary culture. The separation and culture method selects culture conditions of the gonadal tissue of the
tegillarca granosa through a large quantity of basic experiments, so that gonadal cells can be separated, purified, cultured and survive
in vitro.