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Triploid crucian carp tail fin cell line 3nFC as well as construction method and application thereof

A construction method, triploid technology, applied in the direction of artificial cell constructs, animal cells, vertebrate cells, etc., can solve the problems of lack of in vitro cell lines, etc., and achieve the effects of stable cell proliferation, optimized soaking time, and correct number

Inactive Publication Date: 2018-01-30
HUNAN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a triploid Xiangyun crucian carp caudal fin cell line 3nFC and its construction method and application in view of the lack of suitable in vitro cell lines for the current research on the disease resistance mechanism of triploid fish

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  • Triploid crucian carp tail fin cell line 3nFC as well as construction method and application thereof
  • Triploid crucian carp tail fin cell line 3nFC as well as construction method and application thereof
  • Triploid crucian carp tail fin cell line 3nFC as well as construction method and application thereof

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Embodiment

[0041] A triploid Xiangyun crucian carp caudal fin cell line 3nFC of the present invention is preserved in the China Center for Type Culture Collection, and the preservation number is: C201732. The construction method of the triploid Xiangyun crucian carp caudal fin cell line 3nFC is as follows:

[0042] (1) Caudal fin sampling: take 3-9 months old, healthy and fresh triploid Xiangyun crucian carp caudal fins, disinfect them with alcohol with a volume concentration of 75%, put them in a biological safety cabinet, and cut the caudal fins Tissues, soaked in alcohol with a volume concentration of 75% for 40s, rinsed with phosphate buffer for 3 times, and cut into 0.5-1mm 2 small pieces of caudal fin tissue;

[0043] (2) Primary culture: Collect the caudal fin tissue fragments obtained after step (1) and place them in a 1.5mL EP tube, add 1mL of phosphate buffer for rinsing, centrifuge at 1000rpm for 3min, remove the supernatant, Then wash once with 1mL phosphate buffer solution...

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Abstract

The invention discloses a triploid crucian carp tail fin cell line 3nFC which is deposited in China Center for Type Culture Collection, with preservation number CCTCC NO: C201732, the cell morphologyis typical fibroblast cells, the mode of chromosome number is as follows: 3n = 136, the number of chromosomes is correct, and cryopreservation and cell recovery can be carried out; and at the same time the cell line can be transfected and used for gene function studies, so that the foundation for immunology and virology studies for triploid crucian carp is laid, and a solid platform for n vitro studies of triploid crucian carp functional genes is laid. The invention also provides a construction method of the triploid crucian carp tail fin cell line 3nFC. In the method, during primary culture,small tail fin tissue blocks are soaked in fetal bovine serum before drying-sticking, and the soaking time is optimized, so that the small tail fin tissue can relatively well maintain the activity ofcells in the subsequent drying-sticking process after fully absorbing fetal bovine serum, and the emigration time required for the cells is greatly reduced.

Description

technical field [0001] The invention belongs to the field of fish development, and in particular relates to a triploid Xiangyun crucian carp caudal fin cell line 3nFC and its construction method and application. Background technique [0002] The Fish Developmental Biology Laboratory of the College of Life Sciences, Hunan Normal University crossed red crucian carp (Carassius auratasred. Some fertile offspring were found in F1). The diploid offspring F2 was obtained by selfing of F1; F3 was obtained by selfing of F2. F3 is an allotetraploid with both sexes fertile and 200 chromosomes. The allotetraploid crucian carp population is male and female fertile and can stably produce diploid gametes, so that tetraploid characteristics can be continuously and stably inherited. Using the allotetraploid crucian carp as the male parent and the diploid Japanese white crucian carp as the female parent, a triploid Xiangyun crucian carp with 150 chromosomes (3n=150) was prepared by interpl...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 冯浩刘少军傅永明肖军肖俊周维彭灵芝吴慧
Owner HUNAN NORMAL UNIVERSITY
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