Sperm cryopreservation liquid and application in sperm cryopreservation of bostrichthys sinensis

A technology for cryopreservation of Chinese snakeheads, which is applied in the biological field, can solve the problems of unfavorable large-scale production and reproduction of Chinese snakeheads, and poor results, and achieve high sperm motility and in vitro fertilization rate, less damage, and in vitro fertilization rate high effect

Pending Publication Date: 2021-02-12
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when the existing seawater fish sperm cryopreservation method is applied to the sperm cryopreservation of Channa sinensis, the effect is poor, which is not conducive to the large-scale production and reproduction of Channa sinensis

Method used

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  • Sperm cryopreservation liquid and application in sperm cryopreservation of bostrichthys sinensis
  • Sperm cryopreservation liquid and application in sperm cryopreservation of bostrichthys sinensis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 A kind of sperm cryopreservation solution

[0035] The sperm cryopreservation solution includes sperm diluent and antifreeze protection agent. Wherein, the sperm diluent includes the following mass concentrations of components:

[0036] 7.66g / L sodium chloride, 0.51g / L sodium dihydrogen phosphate, 1.16g / L sodium bicarbonate, 0.38g / L potassium chloride, 0.31g / L calcium chloride dihydrate, 1.00g / L D-glucose , 0.26g / L magnesium sulfate heptahydrate.

[0037] The pH of the sperm diluent is 7.0, and the osmotic pressure is 320 mOsmol / kg.

[0038] Cryoprotectants include DMSO, BSA and trehalose.

[0039] When the antifreeze protection agent was added to the sperm diluent, the volume percentage concentration of DMSO was 10%, the mass percentage concentration of BSA was 10%, and the mass concentration of trehalose was 38.7g / L.

Embodiment 2

[0040] Embodiment 2 A kind of sperm cryopreservation solution

[0041] The sperm cryopreservation solution includes sperm diluent and antifreeze protection agent. Wherein, the sperm diluent includes the following mass concentrations of components:

[0042] 7.10g / L sodium chloride, 0.30g / L sodium dihydrogen phosphate, 1.05g / L sodium bicarbonate, 0.20g / L potassium chloride, 0.15g / L calcium chloride dihydrate, 0.50g / L D-glucose , 0.15g / L magnesium sulfate heptahydrate.

[0043] The pH of the sperm diluent is 6.5, and the osmotic pressure is 300 mOsmol / kg.

[0044] Cryoprotectants include DMSO, BSA and trehalose.

[0045] When the antifreeze protection agent is added to the sperm diluent, the mass percent concentration of DMSO is 5%, the mass percent concentration of BSA is 5%, and the mass concentration of trehalose is 35.0g / L.

Embodiment 3

[0046] Embodiment 3 A kind of sperm cryopreservation solution

[0047] The sperm cryopreservation solution includes sperm diluent and antifreeze protection agent. Wherein, the sperm diluent includes the following mass concentrations of components:

[0048] 8.10g / L sodium chloride, 0.70g / L sodium dihydrogen phosphate, 1.30g / L sodium bicarbonate, 0.60g / L potassium chloride, 0.45g / L calcium chloride dihydrate, 1.50g / L D-glucose , 0.35g / L magnesium sulfate heptahydrate.

[0049] The pH of the sperm diluent is 7.5, and the osmotic pressure is 350 mOsmol / kg.

[0050] Cryoprotectants include DMSO, BSA and trehalose.

[0051] When the antifreeze protection agent is added to the sperm diluent, the mass percent concentration of DMSO is 15%, the mass percent concentration of BSA is 15%, and the mass concentration of trehalose is 45.0g / L.

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a sperm cryopreservation liquid and application in sperm cryopreservation of bostrichthys sinensis. Thesperm cryopreservation liquid comprises asperm diluent and an anti-freezing protective agent, wherein the sperm diluent comprises sodium chloride, sodium dihydrogen phosphate, sodium bicarbonate, potassium chloride, calcium chloride dihydrate, D-glucose and magnesium sulfate heptahydrate, and the anti-freezing protective agent comprises dimethyl sulfoxide, bovine serum albumin and trehalose, and the sperm cryopreservation liquid is used for cryopreserving bostrichthys sinensis sperms and is small in damage. The invention further provides a cryopreservation method of bostrichthys sinensis sperms, wherein the method is simple, convenient and rapid, is used for cryopreservation of the bostrichthys sinensis sperms, is high in sperm motility after recovery and high in in-vitro fertilization rate, and can be used for protection of bostrichthys sinensis germplasm resources and large-scale artificial propagation and production.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a sperm cryopreservation solution and its application in cryopreservation of the sperm of Channa sinensis. Background technique [0002] Germ cells produce monoligand sperm and eggs through meiosis, and the sperm and eggs combine to form fertilized eggs, thus producing new individuals. Therefore, germ cells are the basis of population reproduction and important germplasm resources. Germ cell cryopreservation is an effective method for long-term preservation of germplasm resources. Mature fish eggs contain a large amount of lecithin and protein, which makes it extremely difficult to freeze eggs; while fish sperm are small in size, large in number, and relatively simple in content, so sperm is the first choice for fish germ cell cryopreservation. Sperm cryopreservation is of great significance for overcoming the time and geographical constraints of important aquaculture b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 刘威张洪董梦丹张弯弯贾坤同易梅生
Owner SUN YAT SEN UNIV
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