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Composite anti-freezing liquid, and method for preserving human amnion by using same

An antifreeze, human amniotic membrane technology, applied in the application, preservation of human or animal body, animal husbandry, etc., can solve problems such as cell damage, achieve the effect of reducing damage, good permeability, and avoiding ice crystal formation

Inactive Publication Date: 2012-11-07
童亚林
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But antifreezes (especially high concentrations of penetrating antifreezes) can themselves cause cell damage

Method used

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  • Composite anti-freezing liquid, and method for preserving human amnion by using same
  • Composite anti-freezing liquid, and method for preserving human amnion by using same
  • Composite anti-freezing liquid, and method for preserving human amnion by using same

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Experimental program
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Effect test

Embodiment 1

[0041] Embodiment 1: Composite antifreeze

[0042] Take DMEM culture solution, glycerol and dimethyl sulfoxide at a volume ratio of 5:4:1, mix them evenly to obtain a basic antifreeze solution, and then add trehalose to the basic antifreeze solution to control the presence of trehalose in the compound antifreeze solution. The concentration in the frozen solution is 0.1mol / L, and the compound antifreeze solution with a final trehalose concentration of 0.1mol / L is obtained.

Embodiment 2

[0043] Embodiment 2: Composite antifreeze

[0044] Measure DMEM culture solution, glycerin and dimethyl sulfoxide according to the volume ratio of 4.5:4.5:1, mix them evenly to obtain the basic antifreeze solution, then add trehalose to the basic antifreeze solution to control the concentration of trehalose in the compound antifreeze The concentration in the frozen solution is 0.1mol / L, and the compound antifreeze solution with a final trehalose concentration of 0.1mol / L is obtained.

Embodiment 3

[0045] Embodiment 3: Composite antifreeze

[0046] According to the volume ratio of 5.5:3.5:1.5, take DMEM culture solution, glycerol and dimethyl sulfoxide, mix them evenly to obtain the basic antifreeze solution, and then add trehalose to the basic antifreeze solution to control the concentration of trehalose in the compound antifreeze solution. The concentration in the frozen liquid is 0.05 mol / L, and the compound antifreeze liquid with the final trehalose concentration of 0.05 mol / L is obtained.

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Abstract

The invention discloses a composite anti-freezing liquid, and a method for preserving human amnion by using the same. The composite anti-freezing liquid comprises a basic anti-freezing liquid composed of DMEM culture medium, glycerol and dimethyl sulfoxide at volume ratio of (4.5-5.5):(3.5-4.5):(0.5-1.5); and trehalose having a concentration of 0.05-0.15 mol / L in the composite anti-freezing liquid. After pretreatment by the composite anti-freezing liquid, fresh human amnion can be directly preserved in liquid nitrogen without refrigerating with programmed cooling instrument, so as to simplify procedure, save cost, shorten pretreatment time, reduce pollution, and maintain amnion activity in long term.

Description

technical field [0001] The invention relates to the technical field of tissue cell preservation, in particular to a compound antifreeze solution and its application in liquid nitrogen preservation of human amniotic membrane and a method for using the antifreeze solution to preserve human amniotic membrane. Background technique [0002] Human amnion (hereinafter referred to as amnion) is divided into epithelial layer, basement membrane layer, compact layer, fibroblast layer and spongy layer, and the last three layers are also called stroma layer. There are cells and matrix in the amniotic membrane, and the cells are divided into amniotic epithelial cells (stem cells, distributed in the epithelial layer) and amniotic mesenchymal stem cells (distributed in the stromal layer). Stem cell characteristics of different tissue cell differentiation and secretion of various cytokines. Its matrix is ​​also a good cell scaffold material. In addition, amniotic membrane also has the adva...

Claims

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Application Information

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IPC IPC(8): A01N1/02
Inventor 童亚林朱富军詹球杨文贤吕礼芳辛海明朱金红冯小艳
Owner 童亚林
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