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Bovine testis tissue serum-free cryopreservation protective agent for bovine testis tissues, and cryopreservation method and application thereof

A cryopreservation protection agent and cryopreservation method technology, applied in the field of biological cryopreservation, can solve the problems of unfavorable downstream product purification, complex protein composition, exogenous pathogens and viruses, etc., achieve complete testicular tissue structure, reduce cell apoptosis, The effect of maintaining tissue shape

Pending Publication Date: 2020-11-13
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Among them, dimethyl sulfoxide combined with fetal bovine serum (mostly 10% dimethyl sulfoxide and 90% fetal bovine serum) are mostly used in the cryopreservation solution of traditional testicular tissue structure, but due to the protein content in fetal bovine serum Complex, containing harmful substances such as endotoxin and hemoglobin, which is not conducive to the purification of downstream products, it is difficult to maintain consistent quality between production batches, and there is a risk of exogenous pathogens and virus contamination

Method used

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  • Bovine testis tissue serum-free cryopreservation protective agent for bovine testis tissues, and cryopreservation method and application thereof
  • Bovine testis tissue serum-free cryopreservation protective agent for bovine testis tissues, and cryopreservation method and application thereof
  • Bovine testis tissue serum-free cryopreservation protective agent for bovine testis tissues, and cryopreservation method and application thereof

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Embodiment 1

[0028] This embodiment provides a serum-free cryopreservation agent for bovine testicular tissue, which is mixed by 75mL of Knockout DMEM medium, 10mL of KSR serum substitute, 10mL of dimethyl sulfoxide, and 5mL of penicillin and streptomycin obtained and stored at 4°C. Among them, Knockout DMEM medium, KSR serum substitute and penicillin streptomycin can all be commercially available from GIBCO, USA, and dimethyl sulfoxide can be commercially available from Sigma, USA. In addition, among penicillin and streptomycin, the concentration of penicillin is 10000 Units / mL, and the concentration of streptomycin is 10000 μg / mL.

Embodiment 2

[0030] This embodiment provides a serum-free cryopreservation agent for bovine testicular tissue, which is mixed by 62 mL of Knockout DMEM medium, 15 mL of KSR serum substitute, 15 mL of dimethyl sulfoxide, and 8 mL of penicillin and streptomycin obtained and stored at 4°C. Among them, Knockout DMEM medium, KSR serum substitute and penicillin streptomycin can all be commercially available from GIBCO, USA, and dimethyl sulfoxide can be commercially available from Sigma, USA. In addition, among penicillin and streptomycin, the concentration of penicillin is 10000 Units / mL, and the concentration of streptomycin is 10000 μg / mL.

Embodiment 3

[0032] This embodiment provides a serum-free cryoprotectant of bovine testicular tissue, which is mixed by 88mL of Knockout DMEM medium, 5mL of KSR serum substitute, 5mL of dimethyl sulfoxide, and 2mL of penicillin streptomycin obtained and stored at 4°C. Among them, Knockout DMEM medium, KSR serum substitute and penicillin streptomycin can all be commercially available from GIBCO, USA, and dimethyl sulfoxide can be commercially available from Sigma, USA. In addition, among penicillin and streptomycin, the concentration of penicillin is 10000 Units / mL, and the concentration of streptomycin is 10000 μg / mL.

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Abstract

The invention is suitable for the technical field of biological cryopreservation, and provides a bovine testis tissue serum-free cryopreservation protective agent and a cryopreservation method and application thereof. The serum-free cryopreservation protective agent comprises the following components: a Knockout DMEM medium, a KSR serum substitute, dimethyl sulfoxide and mycillin. According to theinvention, the Knockout DMEM culture medium which does not contain serum is used as a substrate, the kSR serum substitute, dimethyl sulfoxide and mycillin are used as low-temperature protective agents in a combined manner, the cost of the cryopreservation protective agent can be reduced, the risk of exogenous pathogen and virus pollution is avoided, in addition, the bovine testis tissue can be preserved at low temperature for a long time, cell apoptosis is reduced, the tissue form is maintained, spermatogenic cells and testis cells are well protected, the testis tissue after cryopreservationand resuscitation is complete in structure, the activity of various testis cells is high, and normal proliferation and development can be achieved.

Description

technical field [0001] The invention belongs to the technical field of biological cryopreservation, and in particular relates to a serum-free cryopreservation protectant for bovine testicular tissue, a cryopreservation method and application. Background technique [0002] The preservation of animal germplasm resources generally includes living species preservation, semen cryopreservation and embryo preservation. Most of the living conservation breeding farms mainly focus on closed breeding, and the entire herd retains the species, which is easily restricted by factors such as group size and market development. In addition, there are problems such as the small number of basic breeding herds and the difficulty in updating and expanding bloodlines of imported or local excellent breeding stock. There are still many difficulties in the work of living species conservation. Although frozen semen technology has been used in production practice, it is not suitable for some newborn o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0215A01N1/0221
Inventor 张学明朱文倩姜禹蔡宁宁杨蕊石建忠
Owner JILIN UNIV
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