Primary culture method of embryonic cells of Tenebrio molitor

An embryonic cell, primary culture technology, applied in animal cells, invertebrate cells, etc., to achieve the effect of reducing labor intensity, strong proliferation ability, and good repeatability

Inactive Publication Date: 2017-07-25
NORTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the in vitro culture of Tenebrio molitor cells is l

Method used

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  • Primary culture method of embryonic cells of Tenebrio molitor
  • Primary culture method of embryonic cells of Tenebrio molitor
  • Primary culture method of embryonic cells of Tenebrio molitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 A kind of primary culture method of Tenebrio molitor embryonic cells comprises the following steps:

[0025] ⑴Collection of Tenebrio molitor eggs: Place Tenebrio molitor adults on a 10-mesh sieve, cover the bottom of the sieve with feed, make the feed contact with the bottom of the sieve, and lay eggs in the dark at 25°C~28°C; The 18-mesh sieve in the feed is used to sieve the eggs, and then remove the impurities in the eggs; incubate for 3 days under the condition of 25°C~28°C, and obtain the hatched Tenebrio molitor eggs, such as figure 1 .

[0026] Among them: Feed refers to the mixture of corn flour and flour at a mass ratio (kg / kg) of 3:2.

[0027] (2) Sterilization of Tenebrio molitor eggs: Under sterile conditions, wrap 1000 grains of Tenebrio molitor eggs in double-layer sterile gauze after full incubation, first soak in 10% NaClO solution for disinfection for 6 minutes, and then soak Disinfect in alcohol with a volume concentration of 75% for 5 m...

Embodiment 2

[0035] Embodiment 2 A kind of primary culture method of Tenebrio molitor embryonic cells comprises the following steps:

[0036] ⑴Collection of Tenebrio molitor eggs: Place Tenebrio molitor adults on a 10-mesh sieve, cover the bottom of the sieve with feed, make the feed contact with the bottom of the sieve, and lay eggs in the dark at 25°C~28°C; Sieve eggs with 18-mesh sieve in the feed, and then remove impurities in the eggs; incubate for 4 days under the condition of 25°C~28°C, and obtain hatched Tenebrio molitor eggs.

[0037] Among them: Feed refers to the mixture of corn flour and flour at a mass ratio (kg / kg) of 3:2.

[0038] (2) Sterilization of Tenebrio molitor eggs: Under sterile conditions, wrap 1000 grains of Tenebrio molitor eggs in double-layer sterile gauze after full incubation, first soak in 10% NaClO solution for disinfection for 6 minutes, and then soak Disinfect in alcohol with a volume concentration of 75% for 5 minutes, then rinse with sterile water with...

Embodiment 3

[0046] Embodiment 3 A kind of primary culture method of Tenebrio molitor embryonic cells comprises the following steps:

[0047] ⑴Collection of Tenebrio molitor eggs: Place Tenebrio molitor adults on a 10-mesh sieve, cover the bottom of the sieve with feed, make the feed contact with the bottom of the sieve, and lay eggs in the dark at 25°C~28°C; The 18-mesh sieve in the feed is used to sieve the eggs, and then remove the impurities in the eggs; under the condition of 25°C~28°C, incubate for 7 days to obtain the hatched Tenebrio molitor eggs.

[0048] Among them: Feed refers to the mixture of corn flour and flour at a mass ratio (kg / kg) of 3:2.

[0049] (2) Sterilization of Tenebrio molitor eggs: Under sterile conditions, wrap 900 grains of Tenebrio molitor eggs in double-layer sterile gauze after full incubation, first soak in 10% NaClO solution for disinfection for 6 minutes, then soak Disinfect in alcohol with a volume concentration of 75% for 5 minutes, then rinse with st...

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Abstract

The invention relates to a primary culture method of embryonic cells of Tenebrio molitor, comprising the steps of (1) collecting eggs of Tenebrio molitor, to be specific, spawning adults of Tenebrio molitor in the shade, and hatching to obtain hatched eggs of Tenebrio molitor; (2) sterilizing the eggs of Tenebrio molitor, to be specific, sterilizing the full hatched eggs of Tenebrio molitor under aseptic condition, flushing, and rinsing to obtain aseptic eggs; (3) acquiring tissues and cells of embryos of Tenebrio molitor, to be specific, transferring the aseptic eggs under aseptic condition to a cell sieve, placing the cell sieve in a culture dish, adding cell culture liquid, and grinding the eggs to obtain cell filtrate; (4) performing primary culture on embryonic cells of Tenebrio molitor, to be specific, culturing the cell filtrate by standing for 15-40 days to obtain an embryonic cell cluster of Tenebrio molitor growing in wall-attaching manner; (5) subculturing embryonic cells of Tenebrio molitor, to be specific, removing the primary culture liquid, digesting the embryonic cell cluster of Tenebrio molitor, dispersing cells by blow-beating, and culturing by standing for 7-10 days before next subculture. The primary culture method is simple to perform, feasible, and high in cell survival rate.

Description

technical field [0001] The invention relates to the technical field of insect cell culture, in particular to a method for primary culture of Tenebrio molitor embryo cells. Background technique [0002] In recent years, insect cell culture technology has been widely used in insect physiology, insect pathology, insect toxicology, molecular genetics, development, endocrinology and other fields. Since Grace established the Anteraeaeucalypti ovarian cell line in 1962, especially since Smith et al. created the Baculovius Expression Vector System (BEVS), the application of insect cell culture technology has been greatly expanded. At present, most of the insect cell lines established in the world are from Lepidoptera and Diptera, and the cell lines from Coleoptera insects only account for about 3%. Coleoptera insects are widely used and are closely related to human life and production. For example, they are used in the extraction of traditional Chinese medicine, food, nutrients and...

Claims

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Application Information

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IPC IPC(8): C12N5/07
CPCC12N5/0601
Inventor 路婉茹臧荣鑫徐红伟杨具田任瑞
Owner NORTHWEST UNIVERSITY FOR NATIONALITIES
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