In vitro culture of amoebocytes from Tachypleus gigas in Leibovitz culture medium

摘要

The present invention provides a process for large scale in vitro production of amoebocytes of Indian Horseshoe Crab (Tachypleus gigas) from dissected gill flaps of T. gigas, in Leibovitz culture medium concentration (2×L-15), having enhanced generation of amoebocytes, said process comprising the steps of: dissecting gill flaps of T. gigas; washing the gill flaps with an antibiotic solution followed by alcohol; culturing the gill flaps in tissue culture plates of sterile saline on a Rocker platform; culturing further the gill flapsin Leibovitz culture medium (2×L-15); purging the gill flaps with Tween 80 solution; and purging again the gill flaps with horse crab serum, keeping the culture viable for 90 days by feeding with fresh medium at an interval of 10-15 days to enable the enhanced release of amoebocytes both within and outside the gill flaps.