Endometrial stem cell and exosome combined preparation for treating endometrial injury and premature ovarian failure, and preparation method and administration method of endometrial stem cell and exosome combined preparation

A technology for endometrial and premature ovarian failure, applied in artificial cell constructs, animal cells, vertebrate cells, etc., can solve the problems of difficult cells and limited therapeutic effect, and achieve the promotion of angiogenesis, maintenance of activity and function, and improvement of local effect of blood supply

Pending Publication Date: 2019-06-18
天津欣普赛尔生物医药科技有限公司
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Problems solved by technology

However, direct intravenous infusion of umbilical cord endometrial stem cells h

Method used

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Examples

Experimental program
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Effect test

preparation example Construction

[0017] A method for preparing a combined preparation of endometrial stem cells and exosomes for treating endometrial injury and premature ovarian failure of the present invention comprises the following steps:

[0018] (1) Cultivation of human endometrial stem cells: select human endometrial stem cells with good growth status and negative microbial detection, P2-P8 human endometrial stem cells, according to 3-5*10 4 / cm 2 The initial density is inoculated in cell culture flasks or cell culture dishes, and the culture medium is a culture system containing fetal bovine serum with a volume ratio concentration of 5-15% in the basal medium;

[0019] (2) After 2-4 days of inoculation and culture, the cell fusion rate reaches 80-95%, and the cell growth state is good. Discard all the culture supernatant, and wash the cells 2-4 times with medical compound electrolyte injection, and then add an appropriate amount of compound electrolyte Solution, make the solution height reach 2-3mm, ...

Embodiment example 1

[0029] Patient Zhang, 34 years old, had endometrial thickness less than 8mm after curettage and curettage, and was clinically diagnosed as thin endometrium.

[0030] Preparation of joint preparations:

[0031] (1) Cultivation of human endometrial stem cells: select P4 human umbilical cord mesenchymal stem cells with good growth status and negative microbial detection, according to 3*10 4 / cm 2 The initial density is inoculated in 10 bottles of T-175 cell culture flasks, and the culture medium is a culture system containing 10% fetal calf serum in the basal medium;

[0032] (2) After 4 days of inoculation and culture, the cell fusion rate reached 90%, and the cell growth state was good. Absorb all the culture supernatant, and wash the cells with medical compound electrolyte injection for 3 times, then add 50ml compound electrolyte solution, and put it into carbon dioxide culture. starvation culture in the box;

[0033](3) After 24 hours of starvation culture, recover all the...

Embodiment example 2

[0039] Patient Xu, 39 years old, suffered from severe endocrine disorders caused by long-term work pressure. The thickness of the endometrium was less than 8mm, and the clinical diagnosis was thin endometrium.

[0040] Preparation of joint preparations:

[0041] (1) Cultivation of human endometrial stem cells: select P6 human endometrial stem cells with good growth status and negative microbial detection, according to 3*10 4 / cm 2 The initial density is inoculated in 20 bottles of T-175 cell culture flasks, and the culture medium is a culture system containing 5% fetal bovine serum in the basal medium;

[0042] (2) After 4 days of inoculation and culture, the cell fusion rate reached 85%, and the cell growth state was good. Absorb all the culture supernatant, and wash the cells with medical compound electrolyte injection for 3 times, then add 40ml compound electrolyte solution, and put in carbon dioxide culture starvation culture in the box;

[0043] (3) After 24 hours of s...

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Abstract

The invention discloses an endometrial stem cell and exosome combined preparation for treating endometrial injury and premature ovarian failure, and a preparation method and administration method of the endometrial stem cell and exosome combined preparation. The endometrial stem cell and exosome combined preparation comprises endometrial stem cells and an endometrial stem cell and exosome concentrated solution obtained through hunger culture. Through the method of hunger culture of the endometrial stem cells, secretion of bioactive substances of the endometrial stem cells is promoted; the bioactive substances secreted by the endometrial stem cells are recovered in an exosome form, thus the recovery efficiency is greatly improved, and the recovery mode is simpler and more convenient; and through a gradient filtration method of biofilms of different hole diameters, exosomes secreted by the endometrial stem cells are collected and concentrated, the recovery process of the exosomes is simplified, and the purity of the exosomes is improved.

Description

technical field [0001] The invention relates to a combined preparation of endometrial stem cells and exosomes, in particular to a combined preparation, preparation method and administration method of endometrial stem cells and exosomes for treating endometrial injury and premature ovarian failure. Background technique [0002] The endometrial injury is mainly the injury of the endometrial basal layer, which is mainly related to curettage during pregnancy. Compared with the normal endometrium, the endometrial basal layer during pregnancy is loose and more susceptible to damage. Based on my country's national conditions and the increasing rate of induced abortion, the occurrence of endometrial damage cannot be ignored. The endometrial basal layer is damaged, which may lead to damage or loss of endometrial stem cells; at the same time, local infection and aseptic inflammation of the injured endometrium can destroy the niche microenvironment of stem cells, resulting in regenerat...

Claims

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Application Information

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IPC IPC(8): A61K35/48A61P15/00C12N5/071
Inventor 鲁振宇张冰晶韩洪起牟鑫煜徐悦
Owner 天津欣普赛尔生物医药科技有限公司
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