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Giant panda milk cryo-preservation solution and giant panda milk preservation method

A preservation method and cryopreservation technology, which are applied in the field of giant panda milk cryopreservation and the preservation of giant panda milk, can solve the problems of loss of milk cells, unfavorable healthy development of giant panda cubs, etc. Normal growth and development, the effect of alleviating freezing injury

Active Publication Date: 2021-03-19
CHENGDU RES BASE OF GIANT PANDA BREEDING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Subsequent systematic testing of the frozen milk found that the nutritional components of the milk frozen by liquid nitrogen direct injection were better preserved, but the cells in the milk were seriously lost, with only about 6% of the frozen survival rate, which is obviously not conducive to giant pandas The healthy development of cubs, therefore, from the perspective of actual production needs, it is urgent to establish a new cryopreservation method for giant panda milk

Method used

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  • Giant panda milk cryo-preservation solution and giant panda milk preservation method

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Embodiment 1

[0022] In order to develop a suitable cryopreservation solution for giant panda milk, the inventor has carried out a lot of meticulous research.

[0023] Because the current conventional 10% DMSO+20% serum mode has a good advantage in maintaining the cell viability, the inventor's research found that the viability of the cells after thawing can reach 96.32% (see grouping research result table). Grouping 1). The reason why this application does not use this cryopreservation solution is because DMSO has certain toxicity and cannot be directly fed to giant pandas after thawing, so it has no practical application effect.

[0024] After a long period of extensive research, the inventor adopted a variety of different combination methods, and finally carried out multiple experiments. For details, see grouping 2-grouping 17 in the grouping research results table. The base liquid used in the freezing solution is all Milk cell culture medium Low-Glucose DMEM.

[0025] In addition, it ...

Embodiment 2

[0032] The present embodiment also provides a method for preserving giant panda milk, comprising the steps of:

[0033] Step 1. Centrifuge the giant panda milk at 630G for 10 minutes, collect the supernatant as milk nutrients, put it into a cryopreservation tube and put it into liquid nitrogen for storage;

[0034] Step 2, the lower sediment part is the milk cell components, resuspended with PBS containing 5% antibiotics, namely penicillin-streptomycin (Penicillin-Streptomycin) solution, centrifuged at 630G for 10 minutes to wash the cells, and repeated once; by washing, the artificial Bacterial contamination and some sundries during milk collection;

[0035] Step 3, according to 1×10 6 ~10 7 The number of cells / mL was adjusted according to the standard, and then 1mL of giant panda milk cryopreservation solution of group 17 was added, mixed gently and transferred to a 1.8mL cryovial. At this time, the number of milk cells in the cryopreservation tube was 1× 106 ~10 7 Befor...

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Abstract

The invention provides a giant panda milk cryo-preservation solution. A basic solution of the cryo-preservation solution is a milk cell culture solution Low-Glucose DMEM, and comprises the following components in concentration: 1.4 M of natural honey, 0.7 M of trehalose, 5 mM of fructose and 100 [mu]M of resveratrol. The invention also provides a preservation method of the giant panda milk. In theapplication, milk nutritional ingredients and cells are separately preserved, and are respectively unfrozen and uniformly mixed in proportion during use, so that the lactation requirements of young pandas can be met. In order to ensure the activity of giant panda milk cells, a special cryoprotectant is adopted. By means of the novel cell cryo-preservation solution, the activity of unfrozen cellsis 90.27%, and the actual production requirements are completely met.

Description

technical field [0001] The invention relates to a freezing liquid and a preservation method of animal milk, in particular to a giant panda milk freezing liquid and a preservation method of the giant panda milk. Background technique [0002] The giant panda (Ailuropoda melanoleuca) is a rare animal endemic to my country and a flagship species of world nature conservation. Improving the success rate of giant panda breeding and raising young has always been the focus of the protection of captive giant pandas, and it is also a difficult point of research. Giant panda cubs are born prematurely, and their later development is completely dependent on breastfeeding. Therefore, the quality of breast milk is a key factor in determining whether the cubs can survive healthily. The composition of giant panda breast milk is complex and diverse, and has many biological functions. It not only provides nutrients for the growth and development of cubs, but also has immune protection function...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0284
Inventor 王东辉侯蓉刘玉良蔡志刚安俊辉兰景超张皓陈敏李媛何平张瑛
Owner CHENGDU RES BASE OF GIANT PANDA BREEDING
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