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Pseudosciaena crocea brain cell line and establishing method thereof

A method of establishment, the technology of large yellow croaker, applied in the direction of artificial cell constructs, animal cells, vertebrate cells, etc., to achieve good stability, vigorous division, and increase the effect of antibacterial spectrum

Inactive Publication Date: 2014-08-27
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the establishment of large yellow croaker cell lines is only reported by Sun Ai in 2010, and the three cell lines of adult large yellow croaker fin, snout, and spleen have been established, and no other tissue cell lines have been reported.

Method used

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  • Pseudosciaena crocea brain cell line and establishing method thereof
  • Pseudosciaena crocea brain cell line and establishing method thereof
  • Pseudosciaena crocea brain cell line and establishing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The establishment of large yellow croaker brain cell line of the present invention:

[0050] (1) Since the large yellow croaker is a crocodile fish and is extremely sensitive to hypochlorite, sterilized seawater containing double antibodies is used instead of hypochlorous acid disinfectant in the body surface disinfection step. The 7-month-old large yellow croaker fry with a length of 9-11 cm were temporarily raised in sterilized seawater containing double antibodies for 1-2 hours. At the end, 3-4 drops of eugenol were dripped until the fish turned over, the abdomen was upward, and there was no stress behavior to the stimulus. Wipe off the mucus on the surface of the fish with a sterilized gauze piece. Wipe the surface of the fish twice with a cotton ball soaked in 75% alcohol. Move the fish body into the ultra-clean bench, take out the brain tissue with dissecting instruments, and put it into a petri dish with D-hanks added before rinsing for 3-4 times.

[0051] (2)...

Embodiment 2

[0055] Cryopreservation and recovery of the large yellow croaker brain cell line of Example 1.

[0056] (1) Frozen storage: Take a bottle of 75cm 2 The cells of the large yellow croaker brain cell line at the bottom of the culture bottle were vigorously grown. Trypsinization was used to collect the cell pellet after centrifugation. Slowly add 3mL of the prepared cell freezing solution, and gently blow the cells to make them evenly dispersed. In cell freezing solution, use a pipette to transfer the liquid into a cryovial. Store the cryopreservation tubes at 4°C for 1 hour, then place them in an ice box, place the ice box at -80°C for 1 day, and finally take out the cryopreservation tubes and immerse them in liquid nitrogen for long-term freezing.

[0057] (2) Take the cryopreservation tube out of the liquid nitrogen and quickly place it in a water bath with adjusted temperature (40°C). During the process of thawing the cells, shake the cryopreservation tube continuously to mak...

Embodiment 3

[0060] The influence of different passage ratio proliferation curves of the large yellow croaker brain cell line of embodiment 1 and the population doubling time analysis:

[0061] From the large yellow croaker brain 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 (that is, add 2.5mL, 1.6mL, 1mL, 0.5mL cell suspension to the 4 culture flasks, and then make up the total amount of each bottle to 5mL 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.

[0062] Such as Figure 8 As shown, for the large yellow croaker...

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Abstract

The invention discloses a Pseudosciaena crocea brain cell line and its establishing method. The cell line is preserved in China Center for Type Culture Collection on Oct., 31, 2013, and has a preservation registration number of CCTCC NO:C2013156. The Pseudosciaena crocea brain cell line can be serially passed, can provide a large amount of the Pseudosciaena crocea brain cell line, and can be used for researching pathogenic characteristics, vaccines, gene functions and breeding; and the Pseudosciaena crocea brain cell line has excellent properties, is an epithelioid cell and is flatly anomaly triangular and polygonal. The Pseudosciaena crocea brain cell line has the characteristics of strong splitting, short passage time, easy digestion, good adherent rate and starvation resistance. The establishing method has strong repeatability, and the established cell line has a good stability.

Description

technical field [0001] The invention belongs to the technical field of fish cell culture, and in particular relates to a large yellow croaker brain cell line and a method for establishing the same. Background technique [0002] Large yellow croaker (Larimichthys crocea), Osteichthyes, Perciformes, Totoabaidae, Yellow croaker. The meat is fresh and tender, the economic value is high, and the market demand is large. Before the 1970s, wild large yellow croaker resources were relatively abundant, and the average annual catch once reached 120,000 tons. In the early 1970s, due to the overfishing of wild large yellow croaker, especially its overwintering groups, the production of wild large yellow croaker was rapidly exhausted. After the 1980s, due to coastal economic development and environmental pollution, the original spawning grounds in the coastal waters were no longer suitable for large yellow croaker spawning, and the catch of wild large yellow croaker is extremely low. I...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12R1/91
Inventor 王艺磊庄道华张子平李敏
Owner JIMEI UNIV
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