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Primary culture method for chilo suppressalis cells

A technology of primary culture of Chilo suppressalis cells, applied to animal cells, etc., to achieve good culture effect and strong repeatability

Inactive Publication Date: 2014-05-28
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are no reports about the in vitro culture technology of C. borer cells and cell lines of C.

Method used

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  • Primary culture method for chilo suppressalis cells
  • Primary culture method for chilo suppressalis cells
  • Primary culture method for chilo suppressalis cells

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Primary culture of fat body cells of Chilo suppressalis larvae

[0031] Go through the following steps:

[0032] (1) In the aseptic operating table (the following operations are all under sterile conditions, and the instruments used must be sterilized or disinfected) immerse one mature larva of Chilo borer in 70% ethanol solution for 10 minutes , carry out surface disinfection, wash the mature larvae of Chilo stem borer with sterile distilled water for 3 times, blot the mature larvae of Chilo stem borer with sterile filter paper or blow dry the mature larvae of Chilo stem borer with a hair dryer;

[0033] (2) Place the blow-dried mature larvae of Chilo suppressalis in a 70% sterilized dissecting wax dish, fix the head and tail with a dissecting needle, cut the stem borer from the back with sterilized dissecting scissors, and then use the dissecting needle to Fix the insect body, take out the fat body tissue with elbow tweezers, keep it as complete as possible during op...

Embodiment 2

[0038] Primary culture of hemolymphocytes from Chilo suppressalis larvae

[0039] (1) In the aseptic operating table (the following operations are all under aseptic conditions, and the instruments used must be sterilized or disinfected) immerse one mature larva of Chilo borer in 75% ethanol solution for 20 minutes , carry out the surface, wash the mature larvae of Chilo stem borer with sterile distilled water for 5 times, blot the mature larvae of Chilo stem borer with sterile filter paper or blow dry the mature larvae of Chilo stem borer with a hair dryer;

[0040] (2) Place the blow-dried mature larvae of Chilo suppressalis in a 75% sterilized dissecting wax dish, fix the head and tail with a dissecting needle, cut the stem from the back with sterilized dissecting scissors, and pipette with 20 μL Aspirate the hemolymph into a culture bottle filled with HBSS cleaning solution (200U / ml penicillin, 0.2μg / ml streptomycin, 0.5μg / ml amphotericin B), tighten the lid, and let stand ...

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Abstract

The invention discloses a primary culture method for chilo suppressalis cells. The primary culture method for the chilo suppressalis cells includes the following steps: (1) a chilo suppressalis larva is soaked in an ethanol solution, surface sterilization is carried out, and the larva body is cleaned and blow-dried; (2) the larva body is placed in a wax dissection tray, the chilo suppressalis larva is dissected, and a tissue or organ for which a cell line needs to be established is taken out and cleaned; (3) the cleaned tissue or organ is placed in a cell culture bottle preliminarily filled with a cell culture solution, the cell culture bottle with the tissue or organ is put into a cell culture box for culture, and the cell culture solution is supplemented after the tissue or organ becomes adherent overnight; (4) the cell culture solution is pipetted and replaced every few days till the culture bottle is filled with expanded and reproduced cells; (5) the whole cell culture solution with newly-reproduced single cells is pipetted and placed into a new culture bottle, and the two culture bottles are placed into the culture box for culture till the cell line is initially established. The primary culture method for the chilo suppressalis cells provided by the invention facilitates chilo suppressalis researches under an isolated culture condition, so as to provide help for the research and development of chilo suppressalis prevention and control methods.

Description

technical field [0001] The invention relates to the technical field of insect cell culture, in particular to a method for primary culture of stem borer cells. Background technique [0002] With the rapid development of life science, cell engineering has been paid more and more attention in the field of bioengineering technology. Cultivating insect cells as research materials has always been an important aspect of scientific research in cell biology, molecular biology and biochemistry, and insect toxicology. [0003] Since Grace first established the cell line of the celestial moth in 1962, insect cell culture technology has developed rapidly, and more than 800 insect cell lines have been established at present, mainly focusing on Lepidoptera and Diptera (Pan Lizhen et al., 1980,1989 ; Zeng Qingtao et al., 1998), Coleoptera (Lynn et al., 1995; Iwabuchi, 1999; Stiles, 1992), Orthoptera (Heinandez-Crespo et al., 2000), Hymenoptera, Homoptera and Hemiptera Most of them are fro...

Claims

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Application Information

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IPC IPC(8): C12N5/07
Inventor 俞晓平刘光富许益鹏
Owner CHINA JILIANG UNIV
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