Patent ID: 6790659
Filing Date: 2004-09-14
Classification: C12N

Abstract:
A process for large scale in vitro production of amoebocytes from Indian Horseshoe Crab (Tachypleus gigas) (T. gigas), said process comprising the steps of:(a) dissecting gill flaps of T. gigas; (b) washing the gill flaps initially with an antibiotic solution for 10 minutes followed by alcohol; (c) culturing the gill flaps in tissue culture plates of sterile saline on a Rocker platform at a temperature ranging between 23-28Â° C.; (d) culturing further the gill flaps of step (c) in Leibovitz L-15 culture medium at a concentration of 2Ã—; (e) purging the gill flaps in the culture medium with Tween 80 solution; and (f) purging again the gill flaps with horseshoe crab serum, while keeping the gill flaps in the medium 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.