Insect epidermis cell line as well as construction method and use thereof

Abstract

The invention provides an insect epidermal cell line and a construction method. The invention successfully overcomes the difficult problem that the epidermal cell tissue has a high differentiation degree, which is difficult to be cultured in vitro. The steps are that the cell culture solution is prepared firstly; then the insect larvae which enter into the molt cycle are selected for the dissection; the dorsal and abdominal epidermis on the body wall of the worm body is taken and rinsed; the dorsal and abdominal epidermis is minced into tiny tissue fragments which are cultured at 25 degree to 29 degree for the adherence to the wall; 1ml to 2ml cell culture solution is added after 1 hour; 1ml to 2ml cell culture solution is added again after 9 days to 11 days; 1:2 to 1:3 dilution subculture is conducted during the passage of generation every 15 days to 20 days; the serum content in culture solution is decreased to 10 percent to 20 percent at the sixth passage and the growth factor content is reduced to half of the original amount; the serum content in culture solution is decreased to 8 percent to 12 percent at the tenth passage and the growth factor content is deprived from the culture solution; the cell line is successfully constructed. The epidermal cell line which is constructed by the method can be passed continuously; the invention provides a large amount of insect epidermal cells for various biological researches.

Description

technical field [0001] The invention relates to an insect epidermis cell line constructed by using insect epidermis tissue, a construction method and application thereof, belonging to insect cell culture technology. Background technique [0002] Since the establishment of invertebrate in vitro culture technology in the 1960s, more than 500 cell lines have been established from more than 100 kinds of insects, such as Spodoptera frugiperda ovary cell line IPLB-Sf21-AE and its clone Sf9( Vaughn et al., 1977), Trichoplusia Tn-5B1-4 cell line, silkworm BmN cell line, American cotton bollworm HzAM1 ovarian cell line, etc., these insect cell lines are used in recombinant protein expression research, baculovirus insecticide It has played and played an important role in the production and research of the amplification of cells, the induction of apoptosis and the study of its mechanism, and the regulation of gene expression in cells by hormones or their analogs. However, the source t...

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Problems solved by technology

However, the source tissues of these insect cell lines are mainly ovaries, testis, embryos, imaginal discs, blood cells, midgut, fat body, etc., while the cell lines constructed from nervous tissue, epidermal tissue, endocrine system, and muscle tissue are rarely or There is no report, and the reason is mainly related to the high degree of cell differentiation of these tissues, which are harsh on the conditions of in vitro culture, and the insufficient composition of the primary cell culture medium is an important factor that makes it difficult for the cells to survive. Improper site and sampling time are also important factors that make primary cell culture difficult to survive.

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