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Large yellow croaker ovarian tissue cell line and application thereof

A technology of ovarian tissue and large yellow croaker, applied in tissue culture, animal cells, reproductive tract cells, etc., can solve the problems that limit the research on the mechanism of gonad development of large yellow croaker, achieve the effect of improving mitotic activity, good stability, and inhibiting bacterial growth

Active Publication Date: 2021-06-01
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no relevant studies and reports on the cell line of the ovarian tissue of large yellow croaker, which greatly limits the research on the mechanism of gonad development in large yellow croaker

Method used

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  • Large yellow croaker ovarian tissue cell line and application thereof
  • Large yellow croaker ovarian tissue cell line and application thereof
  • Large yellow croaker ovarian tissue cell line and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Establishment of large yellow croaker ovarian tissue cell line

[0055] S1. Since the large yellow croaker is a crocodile fish and is extremely sensitive to hypochlorite, sterilized seawater containing double antibodies was used instead of hypochlorous acid disinfectant in the body surface disinfection step. 7-month-old large yellow croaker juveniles with a length of 9-11 cm were temporarily raised in sterilized seawater containing double antibodies for 1-2 h. At the end, 3-4 drops of eugenol were dripped until the fish turned over, the abdomen was upward, and there was no stress response to the stimulation. Wipe off the mucus on the surface of the fish with a sterilized gauze piece. Wipe the surface of the fish twice with gauze soaked in 75% alcohol. Move the fish body into the ultra-clean bench, take out the ovarian tissue with dissecting instruments, and rinse it in PBS solution containing penicillin, streptomycin and amphotericin B (100mL PBS solution wi...

Embodiment 2

[0059] Example 2 Cryopreservation and recovery of large yellow croaker ovarian tissue cell lines

[0060] S1. Freezing: Take a bottle of 75cm 2 The cells of the large yellow croaker ovary tissue cell line that grew vigorously covered the bottom of the culture bottle were digested with trypsin, and the cell pellet was collected after centrifugation, and 3 mL of the prepared cell freezing solution was slowly added, and the cells were gently blown to make them Evenly disperse in the cell freezing solution, and use a pipette to transfer the liquid into the cryovial. Place the cryopreservation tube at 4°C for 1 hour, then place it in a programmed gradient cooling freezer box (temperature drop rate is 1°C / min), place the freezer box at -80°C for 1 day, and finally take out the cryopreservation tube and dry it. Immersed in liquid nitrogen, can be frozen for a long time.

[0061] S2. Take the cryopreservation tube out of the liquid nitrogen and quickly place it in a water bath with ...

Embodiment 3

[0063] Example 3 Proliferation curves of different passage ratios of large yellow croaker ovarian tissue cell lines

[0064] From the large yellow croaker ovarian tissue cell line established in Example 1, cells with good morphology and vigorous proliferation were selected, digested and passaged, and passaged to 25 cm at 1:2, 1:3, 1:5 and 1:10, respectively. 2 Cell culture flasks (i.e. add 2.5 mL, 1.6 mL, 1 mL, 0.5 mL of cell suspension to the 4 culture flasks, and then make up the total amount of each bottle to 5 mL with culture medium). Photographs were taken every 24 hours from the 24th hour of subculture, and the cultured cells were taken by inverted microscope using the classic five-point cross sampling method to count the number of cells observed in the visual field, and the sum of the number of cells observed at five points in each group was plotted for analysis.

[0065] Such as Figure 14 As shown, for large yellow croaker ovarian tissue cell lines, different subcult...

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Abstract

The invention discloses a large yellow croaker ovarian tissue cell line and application thereof. The large yellow croaker ovarian tissue cell line is a large yellow croaker ovarian tissue cell line LYCO and is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: C202117. The large yellow croaker ovarian tissue cell line is excellent in cell character, mainly composed of long fibrous cells, easy to digest, good in adherence rate and resistant to hunger, and the constructed cell line is good in stability. The blank of research on the large yellow croaker ovarian tissue cell line is filled, and a new research tool is provided for research on the gonad development mechanism of the large yellow croaker.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a large yellow croaker ovarian tissue cell line and application thereof. Background technique [0002] Large yellow croaker (Larimichthys crocea), Osteostichthys, Perciformes, Totoaceae, Yellow croaker, mainly inhabits the middle and lower layers of coastal and offshore waters, is the main economic fish in China's coastal waters, and is one of the traditional "four seafood" in China (large yellow croaker, small yellow croaker, hairtail, squid). At present, the main production areas of large yellow croaker are Fujian, Zhejiang and Guangdong. In 2019, Fujian produced 186,514 tons, accounting for 82.7%; Zhejiang produced 23,932 tons, accounting for 10.6%; Guangdong produced 15,103 tons, accounting for 6.7%. However, factors such as the depletion of large yellow croaker broodstock resources, germplasm degradation, and frequent occurrence of large yellow croaker diseases have greatly re...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2500/84C12N2500/44C12N2500/30C12N2501/115C12N2501/11C12N2501/12C12N2500/80Y02A40/81
Inventor 王艺磊钟照威姜永华张子平赵丽萍
Owner JIMEI UNIV
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