The application discloses a preparation method of silkworm blood
lymph fluid with inhibited melanization reaction and low endotoxin, which comprises the following steps: low endotoxin silkworm variety selection, silkworm body disinfection and
impurity removal, silkworm body low-temperature
anesthesia, blood
lymph fluid collection, anti-melanization treatment,
phenol oxidase inactivation treatment,
sediment removal and anaerobic preservation, so as to finally obtain the silkworm blood
lymph fluid with inhibited melanization reaction and low endotoxin. The silkworm blood
lymph fluid extracted by the prior art contains a large amount of endotoxin, which is extremely harmful to in-vitro
cell culture and is one of the bottlenecks for replacing
fetal bovine serum with
insect blood
lymph fluid. The application can reduce the endotoxin concentration in the silkworm
lymph fluid. In view of the problems caused by
quinone derivatives generated by melanization reaction, the application can reduce the
phenol oxidase proenzyme activity in the silkworm blood lymph fluid, long-term inhibit the melanization reaction of the blood lymph fluid, reduce the accumulation of toxic quinones, and make the prepared silkworm blood lymph fluid capable of being used for long-term in-vitro
cell culture.