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Establishment method for pomacea canaliculata ovary cell line

A technology of ovarian cells and construction methods, which is applied in the field of construction of apple snail ovarian cell lines, can solve the problems of endangering crop production, outbreaks of apple snails, etc., and achieve the effect of strong repeatability and good cultivation effect

Inactive Publication Date: 2016-08-17
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Later, due to the failure of marketization, apple snails were abandoned and spread rapidly, resulting in an outbreak of disasters, seriously endangering crop production (Naylor, 1996)

Method used

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  • Establishment method for pomacea canaliculata ovary cell line
  • Establishment method for pomacea canaliculata ovary cell line
  • Establishment method for pomacea canaliculata ovary cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Preparation of cell primary culture medium (preparation of 500mL):

[0046]

[0047]

[0048] In the aseptic operating table, mix the above ingredients and store at 4°C for later use.

[0049] Preparation of cell subculture medium (preparation of 500mL):

[0050] Ingredient Quantity

[0051] RPMI1640 medium 450mL

[0052] Fetal bovine serum 50.0mL

[0053] In the aseptic operating table, mix the above ingredients and store at 4°C for later use.

Embodiment 2

[0054] Primary culture and subculture of embodiment 2 apple snail ovarian tissue cells

[0055] Go through the following steps:

[0056] (1) prepare cell primary culture medium;

[0057] (2) Select female apple snails with a body weight of 30 grams and raise them in sterile water for 12 hours;

[0058] (3) Submerge the female apple snail in 70% ethanol solution and disinfect the surface for 10 minutes;

[0059] (4) Wash the apple snails 3 times with sterile distilled water;

[0060] (5) blot the surface moisture of the snails with sterile filter paper;

[0061] (6) Place the snails in a dissecting dish sterilized by 75% alcohol, and dissect the apple snails;

[0062] (7) Cut the ovarian tissue with dissecting scissors, put it into a six-well culture plate filled with HBSS cleaning solution and wash it 3 times, and cut it into 1mm with sterile scissors for the last time 3 small tissue blocks;

[0063] (8) Inoculate the treated tissue in (7) in a T-25cm cell containing 1mL...

Embodiment 3

[0067] Primary culture and subculture of embodiment 3 apple snail ovarian tissue cells

[0068] Go through the following steps:

[0069] (1) prepare cell primary culture medium;

[0070] (2) select the apple snail with a body weight of 40 grams and raise it in sterile water for 12 hours;

[0071] (3) Submerge the female apple snail in 75% ethanol solution and disinfect the surface for 15 minutes;

[0072] (4) Wash the apple snails 5 times with sterile distilled water;

[0073] (5) blot the surface moisture of the snails with sterile filter paper;

[0074] (6) Place the snails in a dissecting dish sterilized by 75% alcohol, and dissect the apple snails;

[0075] (7) Cut the ovarian tissue with dissecting scissors, put it into a six-well culture plate filled with HBSS cleaning solution and wash it 5 times, and cut it into 1mm with sterile scissors for the last time 3 small tissue blocks;

[0076] (8) Inoculate the treated tissue in (7) in a T-25cm cell containing 1.5mL cel...

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Abstract

The invention discloses an establishment method for a pomacea canaliculata ovary cell line and belongs to the technical field of freshwater organism cell culture. The establishment method includes the following steps that cell culture fluid is prepared; female pomacea canaliculata with the body weight of 30-40 g is selected to be raised in sterile water; the surface of the female pomacea canaliculata is disinfected; the pomacea canaliculata is dissected to obtain ovarian tissue; the ovarian tissue is cleaned with PBS cleaning fluid and cut into pieces; a cell culture bottle is inoculated with the treated ovarian tissue; the culture bottle is placed in a 5% CO2 culture box, and constant temperature culture is conducted at the temperature of 28 DEG C; after overnight treatment, a proper amount of cell culture fluid is added, and the tissue is immersed; the cell culture fluid is sucked out and replaced with an equal amount of cell culture fluid every other 5 days till the culture bottle is filled with extended and proliferated cells. The pomacea canaliculata ovary tissue cell line can be rapidly and repeatedly established within a short time, needed cell culture equipment is simple, and operability is high. The establishment method is an important supplement for culture of cells of aquatic invertebrates and provides fundamental data for research on the immunologic mechanism of the aquatic invertebrates.

Description

technical field [0001] The invention belongs to the technical field of freshwater biological cell culture, and in particular relates to a method for constructing apple snail ovary cell lines. Background technique [0002] Since the in vitro culture method of animal tissues and cells was created by Harrison and Carrel at the beginning of last century, cell culture technology has been greatly developed, and has played an irreplaceable important role in both basic theoretical research and applied research. However, so far, the research on cell culture technology itself has mainly focused on vertebrates (especially mammals) and insects, and relatively little attention has been paid to the cell culture of aquatic invertebrates. Of the papers published in open journals on cell culture issues, the vast majority are of vertebrates and insects. [0003] Since the 1960s, researchers began to pay attention to the cell culture of aquatic invertebrates. Various tissues and cells of aqua...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/07
CPCC12N5/0601
Inventor 刘光富张鹏军杨倩倩许益鹏申屠旭萍
Owner CHINA JILIANG UNIV
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