Shrinkage-method sequential vitrification refrigerating fluid with impervious protecting agent for oocyte and usage method thereof

Abstract

The invention discloses shrinkage-method sequential vitrification refrigerating fluid with an impervious protecting agent for oocyte and a usage method thereof. The sequential vitrification refrigerating fluid comprises shrinkage fluid, equilibrium fluid and vitrification fluid, wherein the shrinkage fluid is impervious protecting agent for dehydrating a cell. The shrinkage-method sequential vitrification refrigerating fluid has the advantages that (1), before entering the equilibrium fluid, the cell is firstly subjected to pretreatment by using the impervious protecting agent to be dehydrated; the acting time in the equilibrium fluid can be shortened; the chemical toxicity of the pervious protecting agent is decreased; (2), the impervious protecting agent is subjected to the pretreatment, so that the stability of the cytomembrane of an ovum can be improved; the oven is enabled to be more tolerant to an osmotic injury; the improvement of the survival rate of a resurgent oven is facilitated; (3), operators of different proficiency levels can all obtain a quite stable high survival rate; the long-time and large-sample-quantity technical training cannot need to be carried out before clinical application; (4), the result is stable; the operation is simple; the clinical wide popularization of a vitrification refrigeration technique for a human oocyte is greatly facilitated.

Description

technical field [0001] The invention belongs to the field of assisted reproduction in medicine, and more specifically relates to a shrinkage method of an oocyte non-permeable protective agent followed by a vitrification freezing liquid and a use method. Background technique [0002] The value of egg freezing is huge. For example, women of the right age suffering from cancer can freeze their eggs before chemotherapy, and then thaw their eggs after cancer treatment without borrowing eggs; professional women can freeze their eggs when they are young, and thaw their eggs when they want to have children Have children; under the premise of ethical and informed consent, women undergoing assisted reproductive therapy can freeze part of their eggs, and donate the frozen eggs to others after successful delivery, so as to solve the problem of the source of donated eggs; eggs are frozen in rare It also has important research and application value in the fields of animals, experimental a...

AI Technical Summary

Problems solved by technology

However, vitrification technology has high requirements for operators, and the outcome of freezing is severely limited by the operating skills and proficiency of embryologists. There are three manifestations: 1) The literature shows that different laboratories that use the same freezing liquid and freezing method have different results. There are great differences in freezing outcomes; 2) There are great differences between different operators in the same laboratory. Take the widely used clinical 15% EG + 15% DMSO + 0.5M sucrose + Cryotop vitrification solution as an example, the operation The difference in survival rate between operators can be as high as 30% or more; 3) The same operator uses the same freezing method, after training with a large sample size, the survival rate can be significantly improved until it is close to the upper limit of his personal performance

This shows that the currently known human egg vitrification scheme (including freezing liquid and freezing method) is difficult to master, and the fact is also the same. This technology is not widely used clinically, and it is only used as a method of radiotherapy and chemotherapy for azoospermia on the day of egg retrieval or cancer patients. pre-remedial therapy

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