Bovine early embryo trophoblast tissue obtaining method

Abstract

The present invention belongs to the field of biotechnology, and provides a bovine early embryo trophoblast tissue obtaining method, which comprises: (1) obtaining a pregnant cow, wherein a pregnant cow is accurately diagnosed within an oestrus period (at least within 18days) after artificial insemination by combining progesterone detection and fluorescence quantitative PCR detection after oestrusinduction and superovulation treatment is performed on cows; (2) picking the uterus of the pregnant cow, flushing embryos, and obtaining an early embryo, wherein the uterus and the associated tissuestructure of the pregnant cow with a gestation time of 18-44 days are picked, and the uterus horn of the uterus with the well-developed ovary corpus luteum is subjected to embryo flushing to obtain the early embryo; and (3) observing the shape of the early embryo tissue, and separating a trophoblast tissue, wherein the tissue structure of the embryo is observed under a microscope, the intact pregnant tissue is fixed with 4% paraformaldehyde, HE staining is performed, and the trophoblast tissue is determined and is stripped. According to the present invention, the method is highly precise, economic and effective.

Description

technical field [0001] The invention belongs to the field of biotechnology, and more specifically relates to a method for obtaining bovine early embryo trophoblast tissue. Background technique [0002] Embryo implantation is a key step in mammalian reproduction. At the maternal-embryo interface, trophoblast cells and maternal endometrial cells play a decisive role in the process of pregnancy recognition and embryo implantation, especially trophoblast cells, which play a leading role in the process of embryo implantation, and their invasion and proliferation And the disorder of functional regulation is closely related to the occurrence and development of various pregnancy-related diseases. At the same time, trophoblast cells are an important part of the placental barrier, the first barrier for maternal blood to enter the fetus, and play an important role in the identification and establishment of maternal pregnancy immune tolerance. Using a whole set of technical methods to...

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Problems solved by technology

However, the placenta gradually ages with the increase of gestational months, and the trophoblast cells also age accordingly, resulting in a small number of cells, low vitality, and more fibroblasts, mesenchymal cells and other cell components, which are difficult to guarantee. Stable

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