Insect epidermis cell line as well as construction method and use thereof
An epidermal cell and construction method technology, applied in the field of insect cell culture, can solve the problems of insufficient primary cell culture medium components, difficulty in surviving primary cell culture, inappropriate sampling time, etc. Timely and specific effects
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[0024] Example: 1. Construction of Helicoverpa armigera epidermal cell line, the steps are as follows:
[0025] (1) Prepare cell culture fluid: add 3.33g / L of hydrolyzed milk protein, 3.33g / L of yeast extract and 0.5g / L of L-glutamine to Grace's medium, filter by suction, and pack , Frozen at -20℃; when used, add 50IU / mL gentamicin, 100IU / mL insulin, 20% of the total volume of fetal bovine serum, and store at 4℃;
[0026] (2) Primary culture: select the 5th instar larvae of cotton bollworm with bursting head shell that enter the moulting cycle, bathe in crushed ice until unconscious, disinfect the body surface with 75% alcohol, and dissect aseptically, take the dorsal and abdominal epidermis of the middle part of the body wall and use Ophthalmic nickel gently separates and removes non-epidermal tissues such as fat body and tracheal tissue. After rinsing with cell culture solution, scrape the fresh epidermal layer with the blunt end of clock forceps, cut it into small tissue pieces...
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