Method for building external nidation model of embryo body and usage

A method of establishing a method and technology of embryoid bodies, which is applied in the field of reproductive medicine, can solve problems such as the inability to observe and study human embryos, and achieve avoiding excessive population growth, major social and economic benefits, and improving the level of treatment technology and assisted reproductive technology Effect

Inactive Publication Date: 2006-12-06
刘芳
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Problems solved by technology

Therefore, this model can only observe the process of the second stage of embryo implantation
In addition, the implantation model established by the co-culture of human decidual cells and mouse blastocysts can only be observed that mouse blastocysts can hatch, adhere, and spread on human decidua, and human embryos cannot be implanted for the first time. The Observation and Research of Three Stages: Blastocyst Invasion of Endometrium Stromal Cells

Method used

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Embodiment Construction

[0022] 1. Selection of experimental materials: endometrium was collected from women aged 29-40 who underwent hysterectomy or curettage due to benign gynecological diseases. The menstrual cycle of the above-mentioned patients should be regular, and they have not taken any hormone drugs in the past three months. Embryos are donated by patients undergoing IVF treatment.

[0023] 2. Isolation and culture of human endometrial stromal cells and induction of decidualization:

[0024] ①. Cleaning: Wash the aseptically collected endometrial tissue with DMEM culture medium for 3 to 5 times to remove blood and mucus.

[0025] ②. Preparation of particles: cut the endometrial tissue collected above into 1mm 3 sized particles for easier digestion.

[0026] ③. Digestion: Put the above-mentioned endometrial tissue particles into a centrifuge tube with a volume of 30ml and add 10-20ml of collagenase I with a concentration of 330IU / ml. Digest with shaking in a water bath for 60 minutes. Whe...

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Abstract

The invention discloses a method for establishing an embryo in vitro implantation model and the user thereof, and the method comprises the courses of: endometrium selection, separate culture and deciduate induction of substrate cells of human endometrium and, embryo culture, co-culture of human embryo and substrate cells of deciduate endometrium, etc, and because it establishes a model of in vitro co-culturing substrate cells of deciduate human endometrium and human embryo and has observed the whole implantation course of human embryo, including adhesion, epitaxial growth and invasion. And the model is nearer to human embryo in vitro implantation environment, and can be used to make a series of researches on related influence factors of embryo implantation, implements in vitro observation of human embryo implantation and provides conditions for researching human embryo implantation mechanism. And the invention can improve sterility curing level and auxiliary procreation technical level, and has great social and economical benefits.

Description

technical field [0001] The invention relates to the field of reproductive medicine, in particular to a method for establishing a human embryo in vitro implantation model and its application. Background technique [0002] At present, with the improvement of infertility treatment technology, especially the development of assisted reproductive technology, the acquisition rate of mature oocytes, the success rate of in vitro fertilization and the implantation rate of embryos have all increased significantly, but the pregnancy rate is very low. The reason is the failure of embryo implantation. Embryo implantation refers to the process in which the blastocyst gradually embeds into the endometrium. It can be divided into three stages: positioning / adhesion, breakthrough and invasion. The process is complex and delicate. In the past, due to the limited means of clinical observation and the difficulty in obtaining endometrium after pregnancy, the implantation process of human embryos ...

Claims

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Application Information

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IPC IPC(8): G09B23/28C12N5/08C12N5/073
Inventor 刘芳高亚萍
Owner 刘芳
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