A method for establishing insect cell lines
An insect cell line and insect cell technology, applied in the direction of microorganism-based methods, animal cells, invertebrate cells, etc., can solve the problems of time-consuming and labor-intensive, low success rate, etc., and achieve strong operability and high cell transformation efficiency Effect
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Embodiment 1
[0051] The establishment of embodiment 1 beet armyworm pupal ovary cell line
[0052] 1) cultivating insect tissues in insect cell culture fluid to obtain dispersed primary insect cells;
[0053] 1.1 Get the female pupae of beet armyworm (from the Institute of Zoology, Chinese Academy of Sciences) the beet armyworm larvae are raised under the conditions of 26 ± 1.5% ℃ and relative humidity of 65%-75% using artificial diet, and submerged in the ultra-clean workbench. 3% sodium hypochlorite solution for 5 minutes, and 75% ethanol solution for 15 minutes to carry out surface disinfection, then clean the insects with sterile distilled water, and then blot dry the surface moisture of the pupae.
[0054] 1.2 Dissect and remove the ovarian tissue, and try to keep it intact during the operation.
[0055] 1.3 Wash the tissue with Ringre's normal saline for 2-3 times, and then use insect cell culture medium (based on Insect-Xpress, containing 100U / mL penicillin, 100U / mL streptomycin ...
Embodiment 2
[0059] Example 2 Establishment of Spodoptera litura pupal ovary cell line
[0060] 1) cultivating insect tissues in insect cell culture fluid to obtain dispersed primary insect cells;
[0061] 1.1 The female pupae of Spodoptera litura (from the Institute of Zoology, Chinese Academy of Sciences) were taken, and the larvae of Spodoptera litura were reared under the conditions of 26±1.5%°C and relative humidity of 65%-75% using artificial diet. Submerge in 3% sodium hypochlorite solution for 5 minutes and 75% ethanol solution for 15 minutes in the ultra-clean workbench to disinfect the surface of the worm body.
[0062] 1.2 Dissect and remove the ovarian tissue, and try to keep it intact during the operation.
[0063] 1.3 Wash the tissue with Ringre's normal saline for 2-3 times, and then use cell culture medium (based on Insect-Xpress, containing 100U / mL penicillin, 100U / mL streptomycin and 10% (v / v) fetal Bovine serum, pH=6.5) was washed once, and placed in a 25cm 2 Adhesi...
Embodiment 3
[0067] Embodiment 3 establishment of beet armyworm pupal ovary cell line
[0068] 1) cultivating insect tissues in insect cell culture fluid to obtain dispersed primary insect cells;
[0069] 1.1 Get the male pupae of beet armyworm (from the Institute of Zoology, Chinese Academy of Sciences) the beet armyworm larvae use artificial feed at 26 ± 1.5% ℃, raise under the condition of relative humidity 65%-75%, immerse in ultra-clean bench 2% sodium hypochlorite solution for 6 minutes, and 75% ethanol solution for 10 minutes to carry out surface disinfection, then clean the insects with sterile distilled water, and then blot the water on the surface of the pupae.
[0070] 1.2 Dissect and remove the testis tissue, and try to keep it intact during operation.
[0071] 1.3 Wash the tissue with Ringre's normal saline for 2-3 times, and then use insect cell culture medium (based on Insect-Xpress, containing 100U / mL penicillin, 100U / mL streptomycin and 10% (v / v) Fetal bovine serum, pH...
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