Method for carrying out embryo in-vitro culture by means of three-dimensional sequential co-culture

Abstract

The invention discloses a method for carrying out embryo in-vitro culture by means of three-dimensional sequential co-culture. The method comprises the following steps of: preparing I type liquid rat tail collagen; separating an oviductal epithelial cell of a rabbit, inoculating the oviductal epithelial cell on the I type liquid rat tail collagen, and culturing the cell to establish a three-dimensional culture system for the oviductal epithelial cell; separating an endometrial epithelial cell of the rabbit, inoculating the endometrial epithelial cell on the I type liquid rat tail collagen, and culturing the cell to establish a three-dimensional culture system for the endometrial epithelial cell; pouring the culture solution in the three-dimensional culture system for the oviductal epithelial cell, replacing the culture solution by a CZB culture solution, and culturing a mouse fertilized egg in the three-dimensional culture system for the oviductal epithelial cell; and pouring the culture solution in the three-dimensional culture system for the endometrial epithelial cell, replacing the culture solution by a CZB culture solution, and transferring a mouse embryo developed to morula stage into the three-dimensional culture system for the endometrial epithelial cell for culturing. The method disclosed by the invention can be used for obviously improving the in-vitro development rate of the mouse embryo and improving the development quality.

Description

technical field [0001] The invention belongs to the field of in vitro culture of embryos, and in particular relates to a method for in vitro culture of embryos by using three-dimensional sequential co-cultivation. Background technique [0002] Embryo culture in vitro has gone through the stages of embryo culture with separate special culture medium, sequential co-culture with different culture medium, co-culture with monolayer helper cells (Helper Cell), and sequential co-culture with different monolayer helper cells. [0003] The study of embryo culture technology in vitro provides the possibility for embryo manipulation in vitro. In vitro embryo culture has greatly promoted the development of disciplines such as breeding, developmental biology, reproductive medicine and embryo engineering. [0004] In recent years, with the continuous improvement of embryo culture medium, the in vitro development rate of embryos has also been greatly improved. However, due to the separat...

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Problems solved by technology

[0006] At present, research on co-cultivation of embryos in vitro is limited to co-cultivation of certain monolayer cultured cells and embryos up to blastocyst. In a closely arranged columnar three-dimensional state, these cells cultured in a single layer in vitro are disorderly arranged in the shape of paving stones, so their morphology and physiological functions are still very different from those in vitro, and cannot well simulate the development environment in vivo

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