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Artificially induced tetraploid crucian cell line and culture method and application thereof

An artificial induction, tetraploid technology, applied in artificial cell constructs, applications, animal cells, etc., can solve the problems of restricting fish polyploid breeding, sterility of hybrid offspring, and long fish sexual maturity. , to achieve the effect of good application prospect, simple process and obvious scientific value

Active Publication Date: 2014-10-22
HUNAN NORMAL UNIVERSITY
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Problems solved by technology

[0003] Hybrid breeding is an important way to breed polyploid fish, but most of the hybrid offspring are sterile or have low fertility. At the same time, fish have a long time to sexual maturity and a long reproductive cycle, which seriously restricts the cultivation of polyploid fish.

Method used

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  • Artificially induced tetraploid crucian cell line and culture method and application thereof
  • Artificially induced tetraploid crucian cell line and culture method and application thereof
  • Artificially induced tetraploid crucian cell line and culture method and application thereof

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Embodiment

[0040] An artificially induced tetraploid crucian carp cell line of the present invention, the artificially induced tetraploid crucian carp cell line is a tetraploid crucian carp cell preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee and the preservation number is CGMCC No.9155 Tie.

[0041] The above-mentioned artificially induced tetraploid crucian carp cell line of the present embodiment is obtained by the following method of in vitro induction of diploid crucian carp cell polyploidy, which specifically includes the following steps:

[0042] 1. Establish a stable diploid crucian carp cell line:

[0043] 1.1 Take the caudal fin treatment: select a young, healthy and vigorous crucian carp, disinfect the caudal fin of the experimental crucian carp with 75% alcohol solution in the preparation room of the cell culture room, and then cut a piece from the tip of the caudal fin with dissecting scissors. Small pieces (abou...

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Abstract

The invention discloses an artificially induced tetraploid crucian cell line which is preserved in the China General Microbiological Culture Collection Center with the preservation number of CGMCC No.9155. The culture method comprises the following steps: by using in vitro cultured diploid crucian cells as a raw material, treating by SP600125 when the cell growth density reaches the degree of fusion being 80-90%; then culturing by a conventional culture medium without SP600125; after culturing a generation, carrying out flow analysis; detecting the content of DNA (Deoxyribonucleic Acid); meanwhile, detecting the chromosome ploidy of the cultured crucian cell; and artificially inducing and sorting and purifying to obtain the in vitro induced tetraploid crucian cell line. The cell nucleus of the tetraploid crucian cell line disclosed by the invention is transplanted to a denucleated egg by using an induced pluripotent stem cell method or adopting a method combining nuclear transplantation. The artificially induced tetraploid crucian cell line can be used for quickly and efficiently creating tetraploid crucian.

Description

technical field [0001] The invention relates to an artificially induced crucian carp cell line and its culture method and application, in particular to an artificially induced tetraploid crucian carp cell line and a method for inducing diploid crucian carp cell polyploidization in vitro and related applications. Background technique [0002] Organisms in nature usually exist in diploid form. However, many organisms have polyploidy events in their evolutionary history. Studies have shown that vertebrates have experienced at least two rounds of tetraploidization. It is currently believed that the formation of new species through chromosome doubling is a feasible evolutionary route. As an important genetic breeding and trait improvement method, polyploid breeding can obtain three or more complete sets of chromosomes, so that the offspring may show variety advantages in terms of growth speed, stress resistance, and disease resistance. , significant economic benefits. [0003]...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N15/873A01K67/027C12R1/91
Inventor 肖亚梅赵小阳周勇华汪妹王姿力
Owner HUNAN NORMAL UNIVERSITY
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