Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Testicle cryopreservation agent for marine fishes and preparation method of spermatogonium

A technology of cryopreservation agent and spermatogonia, applied in the field of marine biology, to achieve the effect of broadening the scope of application, expanding the source of donors, and being simple and easy to operate

Inactive Publication Date: 2017-11-03
MARINE FISHERIES RES INST OF ZHEJIANG
View PDF3 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these studies are mainly aimed at preserving sperm, embryos, tissues, etc., and have not involved whether spermatogonia can be effectively preserved.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Testicle cryopreservation agent for marine fishes and preparation method of spermatogonium
  • Testicle cryopreservation agent for marine fishes and preparation method of spermatogonium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The method for preparing spermatogonial cells utilizing the cryopreservative of the present invention specifically comprises the following steps:

[0031] 1. Prepare 100 mL cryopreservative: 70 mL L-15 culture medium, 10 mL egg yolk, 10 mL 1M trehalose, 10 mL dimethyl sulfoxide (DMSO).

[0032] 2. After anesthetized with MS222, 10 3-month-old (full length: 14.34 ± 0.63 cm; body weight: 34.38 ± 3.98 g) male croakers were anesthetized, and their testes were dissected out.

[0033] 3. Take an appropriate amount of L-15 culture medium in an embryo dish, put fresh testis into it, and cut it into pieces with scissors (the standard size is 3mm×3mm); transfer the cut up testis into a centrifuge tube, centrifuge at 300rpm, and discard. Add L-15 to the supernatant and wash repeatedly 3-5 times.

[0034] 4. Pre-cool the 2.5mL cryovial on ice, and add 2ml of the cryopreservative to it. Transfer the shredded testis into a cryovial with frozen culture medium, and add 5 to 6 pieces ...

Embodiment 2

[0042] 1. Prepare 50 mL cryopreservative: 35 mL L-15 culture medium, 5 mL egg yolk, 5 mL 1M trehalose, 5 mL dimethyl sulfoxide (DMSO).

[0043] 2. After anesthetized with MS222, a 1-year-old (full length: 38.2 cm; body weight: 350.9 g) male squid was anesthetized, and the testes were dissected out.

[0044] 3. Take an appropriate amount of L-15 culture medium in an embryo dish, put fresh testis into it, and cut it into pieces with scissors (the standard size is 3mm×3mm); transfer the cut up testis into a centrifuge tube, centrifuge at 300rpm, and discard. Add L-15 to the supernatant and wash repeatedly 3-5 times.

[0045]4. Pre-cool the 2.5mL cryovial on ice, and add 2ml of the cryopreservative to it. Transfer the shredded testis into a freezing tube with frozen culture medium, and add 5-6 pieces of 3mm×3mm shredded testis to each 2ml of frozen culture medium. Cool on ice for 60 min.

[0046] 5. Move the frozen tubes into the pre-cooled Bicell box, and place them at -80°C f...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a testicle cryopreservation agent for marine fishes and a preparation method of spermatogonium. The preparation method of spermatogonium comprises the following main steps: preparing a cryopreservation agent formula suitable for sciaenops ocellatus; establishing a testicle cryopreservation and unfreezing method; preparing the transplantable spermatogonium. The invention establishes a cryopreservation method for the spermatogonium of the sciaenops ocellatus and technical guarantees are provided for a reproductive cell transplanting technology. According to the testicle cryopreservation agent for the marine fishes and the preparation method of the spermatogonium, a donor source for transplanting reproductive cells is expanded; a testicle is cryopreserved for a long period and can be used according to requirements; guarantees are provided for the donor source for transplanting the reproductive cells and an application range of transplanting of the reproductive cells of fishes is greatly widened.

Description

technical field [0001] The invention belongs to the technical field of marine organisms, and in particular relates to a testis cryopreservation agent of seawater fish and a preparation method of spermatogonia. Background technique [0002] Fish germ cell transplantation, that is, injecting the germ cells of the donor fish into the abdominal cavity of the host fish by microinjection, and the transplanted cells migrate to the gonads through pseudopodia for development, and then form germ cells, and finally produce Mature gametes from donor sources, this technology is also known as the "borrowing" technology of fish, creating a new way for the preservation of germplasm resources of excellent and rare species. After experimental exploration, it was found that type A spermatogonia are effective germ cells for transplantation, which can be extracted and prepared from fish testis. If the fish testis can be frozen for a long time, and the germ cells prepared from the testis after t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 徐冬冬楼宝詹炜陈睿毅刘峰
Owner MARINE FISHERIES RES INST OF ZHEJIANG
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com