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Selection of Mutant Silkworm with Oxidation-deficient Haemolymph for Insect Cell Culture  

Choi, Ji-Young (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
Kim, Jong-Gill (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
Choi, Young-Cheol (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
Yoon, Hyung-Joo (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
Ahn, Mi-Young (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
Kim, Sam-Eun (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
Hwang, Seok-Jo (Department of Agricultural Biology, National Institute of Agricultural Science and Technology, R.D.A.)
Publication Information
Journal of Sericultural and Entomological Science / v.49, no.2, 2007 , pp. 47-50 More about this Journal
Abstract
Insect cell culture system has been demonstrated the effective means of producing medical and agricultural products. Furthermore, Fetal bovine serum (FBS) is in wide use in insect cell culture. Silkworm hemolymph was tested to develop as a substitute for FBS and was effective in insect cell growth. Hemolymph is oxidized and darkens visibly during the collection from silkworms due to the activity of tyrosinase in it. Toxic quinones are produced by the oxidation and consequently inhibit the cell growth. Heat treatment can be used to prevent the oxidation; however, the oxidation may occur during the collection of hemolymph before it is heat-treated. Hemolymphs collected from 257 different strains of silkworms were examined to select the slowly oxidized hemolymphs. Hemolymphs collected from mutant strains such as $Y_4$, TBO and $wE^b$ showed relatively slow color changes. Oxidation rates of the hemolymphs were measured by the absorbance change using a spectrophotometer. The absorbance of mutant hemolymph reached the saturation value at $20^{\circ}C$ in each 330 min ($Y_4$), 360 min (TBO) and 450 min ($wE^b$) min, whereas the total oxidation time of the wild-type (Baekokjam) hemolymph at the same temperature was 120 min. The cell growth in the medium supplemented with mutant species hemolmph was more effective that in the medium supplemented with Baekokjam species hemolymph.
Keywords
Bombyx mori; Hemolymph; Silkworm;
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