Method for culturing and cryopreserving bovine in-vitro fertilization embryos

Abstract

The invention relates to a method for culturing and cryopreserving bovine in-vitro fertilization embryos. The method comprises the following steps of: washing a mature cumulus-oocyte complex in a fertilization culture solution, transferring the mature cumulus-oocyte complex into the fertilization culture solution, and putting the mature cumulus-oocyte complex into an incubator for later use; taking a frozen sperm tubule from liquid nitrogen, unfreezing, taking a sperm suspension, and counting sperms; adding the sperm suspension into fertilization culture solution drops containing oocytes to co-incubate the sperms and the oocytes, completing in-vitro fertilization operation, completely removing granular cells around the embryos by using an oocyte stripping needle, putting the embryos into an embryo culture solution for culturing, transferring available embryos into a freezing solution, and putting the embryos into liquid nitrogen for cryopreservation; and when the cryopreserved embryos need to be used, taking the tubule from the liquid nitrogen, transferring the tubule into an unfreezing solution, putting the embryos into small drops of the embryo culture solution for culturing, and completing the unfreezing process. The invention further relates to the used freezing solution and the used unfreezing solution. According to the method, the survival rate of the bovine in-vitro fertilization embryos after cryopreservation and unfreezing can be remarkably increased.

Description

technical field

[0001] The invention belongs to the technical field of animal reproduction, and relates to a technology for agricultural-animal husbandry and veterinary reproduction, in particular to a method for bovine in vitro fertilization. In addition, the present invention also relates to the use of related working test solutions used in bovine in vitro fertilization. The application in fertilization, in particular, relates to a method of bovine in vitro fertilization, for example, relates to a method of freezing and thawing bovine in vitro fertilization blastocysts with excellent effects. The bovine in vitro fertilization method of the invention has excellent technical effects. Background technique

[0002] In Vitro Fertilization (In Vitro Fertilization) or (external fertilization) refers to the technology in which mammalian sperm and eggs complete the fertilization process in an artificially controlled environment outside the body. It is referred to as IVF in English....