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Method of preparing endometrium mesenchymal stem cells from menstrual blood

A technology of mesenchymal stem cells and endometrium, applied in the field of biomedicine, can solve the problems of cell release, difficulty in use, low proliferation activity of endometrial mesenchymal stem cells, etc., and achieve high proliferation activity and convenient use

Active Publication Date: 2015-12-23
ZHEJIANG SHENGCHUANG PRECISION MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the menstrual cup is inserted into the body for 4-5 hours, taken out for cleaning, disinfection, and then put back into the body, which is not convenient for professional women to use at work
Existing tampons can absorb menstrual blood, but the cells in the menstrual blood absorbed on the tampon cannot be released from the tampon, and endometrial tissue pieces cannot be recovered, so they cannot be used to extract the endometrium mesenchymal stem cells
Therefore, the current method of preparing endometrial mesenchymal stem cells from menstrual blood is difficult to use clinically and the proliferative activity of endometrial mesenchymal stem cells is low.

Method used

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  • Method of preparing endometrium mesenchymal stem cells from menstrual blood
  • Method of preparing endometrium mesenchymal stem cells from menstrual blood
  • Method of preparing endometrium mesenchymal stem cells from menstrual blood

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Embodiment approach

[0036] Wherein, the eluent can be a conventional buffer for various cell cultures, such as phosphate buffer or saline, and according to a preferred embodiment of the present invention, the eluent contains 50-200 μg / mL penicillin, 50-200 μg / mL streptomycin, 1.5-4 μg / mL amphotericin B and 30-50 μg / mL ciprofloxacin and 4-8 mg / mL heparin in buffer at pH 7.2-7.5. In this preferred embodiment, cells in the menstrual blood (including endometrial mesenchymal stem cells) and endometrial tissue pieces adsorbed on the gauze absorbent layer 22 can be more easily eluted from the gauze absorbent layer 22 Down, so as to improve the proliferative activity of endometrial mesenchymal stem cells.

[0037] According to the method of the present invention, the operation of separating nucleated cells from the eluted product may include: mixing the eluted product with erythrocyte lysate, and then centrifuging at (800-1200)×g to remove the liquid, Obtain a precipitate, then cut the precipitate into...

preparation Embodiment 1

[0041] This preparation example is used to illustrate the preparation process of the tampon of the present invention.

[0042] Sterile absorbent cotton gauze (purchased from Guangdong Hengjian Pharmaceutical Co., Ltd., article number is 861587) is cut and then pressed under tampon pressing conditions (pressing method as described in the patent of CN1355684A) to be 6 cm in length and 8 mm in inner diameter. A cylindrical sleeve with a thickness of 3mm, and 8 symmetrical grooves 23 with outward openings are pressed on the gauze absorbent layer 22, and the direction of the grooves 23 is in line with the axial direction of the cylindrical sleeve. Parallel; the maximum depth of the groove 23 is 50% of the maximum thickness of the cylindrical casing, ie 1.5mm. Absorbent cotton (purchased from Beijing Aominsheng Medical Instrument Co., Ltd., article number 001) was filled in the gauze absorbent layer 22 as the absorbent fiber core 24, the average length of the absorbent fiber core 24...

preparation Embodiment 2

[0045] The preparation of the tampon was carried out according to the method of Preparation Example 1, the difference was that before the sterile absorbent cotton gauze was cut and pressed, the coating solution was pretreated; specifically, the sterile absorbent cotton gauze was immersed in the coated Then remove and squeeze out the excess liquid and then dry it. The coating solution contains 50ng / mL polylysine, 1800U / mL heparin and 10ng / μL fibronectin, and the pH Phosphate buffer with a value of 7.4.

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Abstract

The invention provides a method of preparing endometrium mesenchymal stem cells from menstrual blood. The method comprises the steps that firstly, a menstruation tampon is placed in the vagina of a woman during the menstrual period of the woman, wherein the menstruation tampon comprises an absorption tampon body which is provided with a water absorption fiber core, the water absorption fiber core is wrappedina gauze absorption layer, and the menstruation tampon is kept in the vagina till menstrual blood and / or deciduous endometrial tissue are adsorbed on the gauze absorption layer; secondly, a pull wire is pulled out to take out the menstruation tampon, the menstrual blood and / or the deciduous endometrial tissue adsorbed on the gauze absorption layer are eluted off with eluant to obtain an eluted product; thirdly, karyocytes are extracted from the eluted product and subjected to cell culture. The method can be conveniently used clinically and can be accepted by menstrual blood donators; moreover, the obtained endometrium mesenchymal stem cells have higher proliferative activity.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for preparing endometrial mesenchymal stem cells from menstrual blood. Background technique [0002] Mesenchymal stem cells (MesenchymalStemCell, MSC) is a kind of stem cells, named for its ability to differentiate into mesenchymal tissue, has self-renewal ability and multi-directional differentiation potential, and can be used for hematopoietic support and promotion of stem cell implantation and immune regulation, etc. function. There are a considerable number of mesenchymal stem cells in the endometrial tissue shed with women's menstrual blood, which are called endometrial mesenchymal stem cells. The number of endometrial mesenchymal stem cells is large, which is 30 times that of bone marrow-derived stem cells; strong in vitro proliferation ability: doubled in 24 hours, can proliferate 390 times, and can be passaged up to 50 times; high differentiation potential: ...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 余艳春郑俊李程
Owner ZHEJIANG SHENGCHUANG PRECISION MEDICAL TECH CO LTD
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