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Coccidiosis oocyst cryoprotectant, and preparation method and application thereof

A cryopreservation agent and insect egg technology, which is applied in the application, preservation of human or animal bodies, animal husbandry, etc., can solve the problems of increasing the cost of vaccines, time-consuming and labor-intensive, etc., to inhibit the recrystallization process of ice crystals, reduce production costs, and facilitate Achieved effect

Active Publication Date: 2020-09-18
SONGSHAN LAKE MATERIALS LAB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, Eimeria and live coccidia vaccines are stored at 4°C, but the storage period is usually no more than 6 months, and manual subculture intervention is required every two months for the preservation of coccidia vaccines, which is time-consuming and laborious to increase the number of vaccines. cost

Method used

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  • Coccidiosis oocyst cryoprotectant, and preparation method and application thereof
  • Coccidiosis oocyst cryoprotectant, and preparation method and application thereof
  • Coccidiosis oocyst cryoprotectant, and preparation method and application thereof

Examples

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Embodiment 1

[0036]Embodiment 1: This embodiment is illustrated by taking the cryopreservation and recovery of Eimeria maxima oocysts as an example, but it is not used to limit the scope of the present invention.

[0037] (1) Preparation of coccidian oocyst cryopreservation agent: put 90ml of water for injection into a beaker placed in a water bath at 4°C, put in a stirrer, and adjust the rotating speed to 500rpm; add 0.2g of sodium dodecylsulfonate, two Stearyl phosphatidylethanolamine-polyethylene glycol (DSPE-mPEG2000) 0.8g, stirred for 10 minutes; add tartaric acid 5g, ammonium sulfate 2g, sucrose 9g, fructose 5g standard PBS buffer 5ml, sodium dihydrogen phosphate 0.3g and Disodium hydrogen phosphate 0.7g, stirred for 30 minutes.

[0038] (2) pretreatment oocysts, need to carry out following purification pretreatment before adding Eimeria maxima oocysts:

[0039] 1. Take 2ml of Eimeria maxima oocyst fluid and centrifuge (3000rpm, 5min) to precipitate;

[0040] 2. Remove the supernat...

Embodiment 2

[0054] Embodiment 2: This embodiment is basically the same as Embodiment 1, the only difference is that the coccidian oocyst cryopreservation agent comprises the following components in mass percentage: sodium dodecylsulfonate 0.01%, distearoyl Phosphatidylethanolamine-polyethylene glycol 2%, tartaric acid 1%, ammonium sulfate 3%, sucrose 5%, fructose 3%, sodium dihydrogen phosphate 0.5%, disodium hydrogen phosphate 0.8%, standard PBS buffer 4%, remaining The amount is water for injection.

Embodiment 3

[0055] Embodiment 3: The present embodiment is basically the same as Embodiment 1, the only difference is that the coccidian oocyst freezing agent comprises the following components in mass percentage: sodium dodecylsulfonate 0.05%, distearoyl Phosphatidylethanolamine-polyethylene glycol 0.9%, tartaric acid 4%, ammonium sulfate 0.5%, sucrose 10%, fructose 1%, sodium dihydrogen phosphate 0.8%, disodium hydrogen phosphate 0.2%, standard PBS buffer 3%, the rest The amount is water for injection.

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Abstract

The invention discloses a coccidiosis oocyst cryoprotectant, and a preparation method and application thereof. The coccidiosis oocyst cryoprotectant comprises the following components: sodium dodecylsulfate, distearoyl phosphatidyl ethanolamine-polyethylene glycol, tartaric acid, ammonium sulfate, sucrose, fructose, sodium dihydrogen phosphate, disodium hydrogen phosphate, a standard PBS buffer solution and water for injection. The cryoprotectant is reasonable in formula, and can effectively inhibit ice crystal recrystallization in the cryopreservation process, regulate and control ice crystal morphology, reduce ice crystal growth rate and decrease damage of ice crystals to coccidiosis oocysts; and meanwhile, phospholipid molecules can be adsorbed to the walls of the coccidiosis oocysts,so the ice damage resistance of the coccidiosis oocysts is improved, and the coccidiosis oocysts can be stored in liquid nitrogen for a long time. Compared with a traditional cryoprotectant, the coccidiosis oocyst cryoprotectant of the invention does not contain toxic molecules such as DMSO and components such as expensive serum, avoids the toxic damage of DMSO to coccidiosis oocysts, lowers production cost, has effectively improved applicability in the field of coccidiosis vaccines is enhanced, and the cryoprotectant and is beneficial for wide popularization and application.

Description

technical field [0001] The invention relates to the technical field of cryopreservation, in particular to a coccidia oocyst freezing agent, a preparation method thereof and an application in coccidia oocysts. Background technique [0002] In recent years, cryopreservation of cells, tissues and organs has attracted much attention, and breakthroughs in cryopreservation technology will provide strong theoretical and experimental support for the development of regenerative medicine. Active substances that are separated from the maternal environment, such as cells, consume their own nutrients in the cells due to metabolism and other life activities during preservation, and their activity will inevitably weaken with the preservation time until they are completely inactivated. The extremely low temperature, usually 77K (-196°C, the boiling point of liquid nitrogen), can effectively inhibit or even stop life activities such as metabolism in cells, prolong the life of cells and achie...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 王健君赵立山吕健勇
Owner SONGSHAN LAKE MATERIALS LAB
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